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1996 July Powered By Docstoc
					1996 July
From p.sobieszczuk at ic.ac.uk Wed Jul 24 20:01:05 1996
From: p.sobieszczuk at ic.ac.uk (Peter Sobieszczuk)
Subject: test only (disregard)
Message-ID: <v0151010aae1c224e0a34@[155.198.45.54]>

test only (disregard)



From csmits at qlt-pdt.com Thu Jul 25 14:01:16 1996
From: csmits at qlt-pdt.com (Claire Smits)

Subject: Stress indicators of mice left in the dark
Message-ID: <SIMEON.9607251316.A@csmits.qlt-pdt.com>

Hello Transgenic List Subscribers:

Can anyone direct me to any studies, papers, or general information,
regarding stress indicators of mice that have been placed in the
dark or light reduced areas for 24 hours or longer ?

Thank you

Claire Smits
csmits@qlt-pdt.com



From Allison_F._Treloar at ccmail.bms.com Fri Jul 26 09:23:51 1996
From: Allison_F._Treloar at ccmail.bms.com (Allison F. Treloar)

Subject: Contract knock-out production
Message-ID: <9606268383.AA838394810@ccgate0.bms.com>

I hope there are enough of you signed up on this new list to help me
out..

Aside from Genome Systems, does anyone know of other suppliers of
contract
knock-out mouse production services. I don't know if DNX has this
service up
and running yet.

Thanks,

Allison

Technical Supervisor-Transgenics
Bristol-Myers Squibb Co.
Allison_F._Treloar@ccmail.bms.com


From stewarv at cesmtp.ccf.org   Fri Jul 26 11:20:45 1996
From: stewarv at cesmtp.ccf.org (Valerie Stewart M.S.)

Subject: Contract knock-out production -Reply
Message-ID: <s1f89ccd.049@cesmtp.ccf.org>


Allison et. al.--

What do you mean exactly by contract knockout-mouse production? My
facility will accept frozen ES cells for injection, to produce
chimeras for a fee. We do not do the initial production of the ES
cell clones containing the knocked-out gene. We haven't been
breeding for germline transmission either, though we may extend the
service to that. As of yet, we've only had internal business. Let
me know if we can be of assistance.

Valerie Stewart
Transgenic/Knockout Facility
Cleveland Clinic Research Institute
"stewarv@cesmtp.ccf.org"


From r-carver at nimr.mrc.ac.uk Tue Jul 30 10:11:53 1996
From: r-carver at nimr.mrc.ac.uk (Rick Carver)

Subject: Scientific Meeting on Transgenic Animals
Message-ID: <7646.9607290822@nimsn01.nimr.mrc.ac.uk>

Dear All

The Laboratory Animal Science Association (LASA) is establishing a
specialist Scientific Section on Transgenic Animals. This Section will
be
launched with an inaugural meeting entitled "Transgenic Animals-
Progress,
Problems and Potential".

The meeting will be held in London (UK) on Friday 27 September 1996 and
has
the following programme:

Transgenesis- setting the scene
Transgenic animals in Developmental Biology
Transgenic animals in Immunology
Transgenic animals in Therapeutic Protein Production
Transgenic animals in Neuroscience
Transgenic animals in Oncology
Transgenic animals in Pharmaceuticals Discovery and Development
Ethical and legal aspects of the use of transgenic animals

The speakers are from NIMR, ICRF, various University departments and
leading
biotechnology and pharmaceutical companies.

The cost will be 55 pounds stirling for LASA members and 75 pounds
stirling
for non-members. Registration forms are available from:   LASA
Speciality
Sections, PO Box 3993, Tamworth, Staffs, B78 3QU. The closing date for
registrations is 23 Aug 96.




Rick Carver
Head of Biological Services and Institute Veterinarian
National Institute for Medical Research
The Ridgeway
MILL HILL
London                     E-Mail: r-carver@nimr.mrc.ac.uk
NW7 1AA                    Tel: + 44 (0)181 959 3666 ext 2199
United Kingdom             Fax: + 44 (0)181 913 8601

"There is something fascinating about science: one gets such wholesale
returns of conjecture out of such a trifling investment of fact" - Mark
Twain


From TSAUNDER at hg-basic1mail.hg.med.umich.edu Mon Jul 29 12:36:47
1996
From: TSAUNDER at hg-basic1mail.hg.med.umich.edu (TSAUNDER)

Subject: Contract knock-out production
Message-ID: <27CDBA4742@HG-BASIC1MAIL.HG.MED.UMICH.EDU>

Alison,

Lexicon Genetics, Inc. makes contract knock-out mice. Their URL is
http://www.lexgen.com
They are located in Woodlands, Texas (North of Houston). They are
using
Allen Bradley's ES cell line.


Thom Saunders, Ph.D.
Transgenic Animal Model Core
Biomedical Research Core Facilities
University of Michigan Medical School
email: tsaunder@umich.edu


Date:          Fri, 26 Jul 1996 08:23:51 -0500 (EST)
From:          "Allison F. Treloar" <Allison_F._Treloar@ccmail.bms.com>
Subject:       Contract knock-out production
To:            transgenic-list@ic.ac.uk
Reply-to:      transgenic-list@ic.ac.uk

I hope there are enough of you signed up on this new list to help me
out..

Aside from Genome Systems, does anyone know of other suppliers of
contract
knock-out mouse production services. I don't know if DNX has this
service up
and running yet.

Thanks,

Allison

Technical Supervisor-Transgenics
Bristol-Myers Squibb Co.
Allison_F._Treloar@ccmail.bms.com




From p.sobieszczuk at ic.ac.uk Wed Jul 24 19:01:05 1996
From: p.sobieszczuk at ic.ac.uk (Peter Sobieszczuk)

Subject: test only (disregard)
Message-ID: <v0151010aae1c224e0a34@[155.198.45.54]>

test only (disregard)



From csmits at qlt-pdt.com Thu Jul 25 18:01:16 1996
From: csmits at qlt-pdt.com (Claire Smits)

Subject: Stress indicators of mice left in the dark
Message-ID: <SIMEON.9607251316.A@csmits.qlt-pdt.com>

Hello Transgenic List Subscribers:

Can anyone direct me to any studies, papers, or general information,
regarding stress indicators of mice that have been placed in the
dark or light reduced areas for 24 hours or longer ?

Thank you

Claire Smits
csmits@qlt-pdt.com



From Allison_F._Treloar at ccmail.bms.com Fri Jul 26 14:23:51 1996
From: Allison_F._Treloar at ccmail.bms.com (Allison F. Treloar)

Subject: Contract knock-out production
Message-ID: <9606268383.AA838394810@ccgate0.bms.com>

I hope there are enough of you signed up on this new list to help me
out..

Aside from Genome Systems, does anyone know of other suppliers of
contract
knock-out mouse production services. I don't know if DNX has this
service up
and running yet.

Thanks,

Allison

Technical Supervisor-Transgenics
Bristol-Myers Squibb Co.
Allison_F._Treloar@ccmail.bms.com


From stewarv at cesmtp.ccf.org Fri Jul 26 15:20:45 1996
From: stewarv at cesmtp.ccf.org (Valerie Stewart M.S.)
Subject: Contract knock-out production -Reply


Allison et. al.--

What do you mean exactly by contract knockout-mouse production? My
facility will accept frozen ES cells for injection, to produce
chimeras for a fee. We do not do the initial production of the ES
cell clones containing the knocked-out gene. We haven't been
breeding for germline transmission either, though we may extend the
service to that. As of yet, we've only had internal business. Let
me know if we can be of assistance.

Valerie Stewart
Transgenic/Knockout Facility
Cleveland Clinic Research Institute
"stewarv@cesmtp.ccf.org"


From r-carver at nimr.mrc.ac.uk Tue Jul 30 10:11:53 1996
From: r-carver at nimr.mrc.ac.uk (Rick Carver)

Subject: Scientific Meeting on Transgenic Animals
Message-ID: <7646.9607290822@nimsn01.nimr.mrc.ac.uk>

Dear All

The Laboratory Animal Science Association (LASA) is establishing a
specialist Scientific Section on Transgenic Animals. This Section will
be
launched with an inaugural meeting entitled "Transgenic Animals-
Progress,
Problems and Potential".

The meeting will be held in London (UK) on Friday 27 September 1996 and
has
the following programme:

Transgenesis- setting   the scene
Transgenic animals in   Developmental Biology
Transgenic animals in   Immunology
Transgenic animals in   Therapeutic Protein Production
Transgenic animals in   Neuroscience
Transgenic animals in   Oncology
Transgenic animals in Pharmaceuticals Discovery and Development
Ethical and legal aspects of the use of transgenic animals

The speakers are from NIMR, ICRF, various University departments and
leading
biotechnology and pharmaceutical companies.

The cost will be 55 pounds stirling for LASA members and 75 pounds
stirling
for non-members. Registration forms are available from:    LASA
Speciality
Sections, PO Box 3993, Tamworth, Staffs, B78 3QU. The closing date for
registrations is 23 Aug 96.




Rick Carver
Head of Biological Services and Institute Veterinarian
National Institute for Medical Research
The Ridgeway
MILL HILL
London                     E-Mail: r-carver@nimr.mrc.ac.uk
NW7 1AA                    Tel: + 44 (0)181 959 3666 ext 2199
United Kingdom             Fax: + 44 (0)181 913 8601

"There is something fascinating about science: one gets such wholesale
returns of conjecture out of such a trifling investment of fact" - Mark
Twain


From TSAUNDER at hg-basic1mail.hg.med.umich.edu Mon Jul 29 17:36:47
1996
From: TSAUNDER at hg-basic1mail.hg.med.umich.edu (TSAUNDER)

Subject: Contract knock-out production
Message-ID: <27CDBA4742@HG-BASIC1MAIL.HG.MED.UMICH.EDU>

Alison,

Lexicon Genetics, Inc. makes contract knock-out mice. Their URL is
http://www.lexgen.com
They are located in Woodlands, Texas (North of Houston). They are
using
Allen Bradley's ES cell line.


Thom Saunders, Ph.D.
Transgenic Animal Model Core
Biomedical Research Core Facilities
University of Michigan Medical School
email: tsaunder@umich.edu


Date:          Fri, 26 Jul 1996 08:23:51 -0500 (EST)
From:          "Allison F. Treloar" <Allison_F._Treloar@ccmail.bms.com>
Subject:       Contract knock-out production
To:            transgenic-list@ic.ac.uk
Reply-to:      transgenic-list@ic.ac.uk

I hope there are enough of you signed up on this new list to help me
out..

Aside from Genome Systems, does anyone know of other suppliers of
contract
knock-out mouse production services. I don't know if DNX has this
service up
and running yet.

Thanks,

Allison

Technical Supervisor-Transgenics
Bristol-Myers Squibb Co.
Allison_F._Treloar@ccmail.bms.com



1996 August
From Allison_F._Treloar at ccmail.bms.com Fri Aug 2 16:50:53 1996
From: Allison_F._Treloar at ccmail.bms.com (Allison F. Treloar)
Subject: Transgenic Barriers

Hi folks,

I am trying to get a feel for how many institutions are using barrier
conditions
for their transgenic/knockout colonies. Please reply to me off list
and I'll
sumarize for the group. Don't forget to include where you're from!

no/low barrier = conventional caging and access (+/- filter tops on
caging)

modified barrier =   access restrictions, gowning, isolators, (+/-
autoclaving)

full barrier = shower in, isolator caging, autoclaved food and bedding

Thanks in advance!
Allison Treloar
Technical Supervisor - Transgenics
Bristol-Myers Squibb

Allison_F._Treloar@ccmail.bms.com
From mgallardo at wsu.edu Fri Aug 2 14:36:49 1996
From: mgallardo at wsu.edu (Mike Gallardo)

Subject: Transgenic Barriers
Message-ID: <199608022036.NAA14361@cheetah.it.wsu.edu>

At 03:50 PM 8/2/96 -0500, you wrote:
>Hi folks,
>
>I am trying to get a feel for how many institutions are using barrier
conditions
>for their transgenic/knockout colonies. Please reply to me off list
and I'll
>sumarize for the group. Don't forget to include where you're from!
>
>no/low barrier = conventional caging and access (+/- filter tops on
caging)
>
>modified barrier = access restrictions, gowning, isolators, (+/-
autoclaving)
>
>full barrier = shower in, isolator caging, autoclaved food and bedding
>
>Thanks in advance!
>Allison Treloar
>Technical Supervisor - Transgenics
>Bristol-Myers Squibb
>
>Allison_F._Treloar@ccmail.bms.com
>
>
>Here at Wegner Hall Vivarium/WSU we use isolator caging, autoclaved
food
and bedding. We do not shower in, we use steril gowns, booties, mask,
hair
nets and gloves.
>
>

   (''`-''-/").___..--''"`-._
     `o_ o )    `-. (       ).`-.__.`)
     (_Y_.)' ._    ) `._ `. ``-..-'
   _..`--'_..- / /--' _. .'
  (il).-'' (li).'    ((!.-'     Mike Gallardo BS MS LAT
                                Administative Manager

    GO COUGS!!!!               Wegner Hall Vivarium G-44
                               Washington State University
                               Pullman, Wa 99164-6510
                               Phone (509)335-0977
                               Fax   (509)335-0162
                               e-mail mgallardo@wsu.edu
 "Brains are for thinking--pencils are for remembering"
From fmargoli at umabnet.ab.umd.edu Fri Aug 2 18:13:51 1996
From: fmargoli at umabnet.ab.umd.edu (Frank L. Margolis)

Subject: No subject
Message-ID: <v0153050fae282db061cd@[134.192.49.38]>



Anyone-

Is there an up-to-date master list of knockout and/or transgenics that
is
maintained anywhere?

Thanks

Frank L. Margolis Ph.D.
Department of Anatomy
University of Maryland at Baltimore
School of Medicine
685 West Baltimore Street
Baltimore MD 21201

Phone 410-706-8913 office
             -8914 lab
FAX          -2512 Department office




From ignelzi at umich.edu Fri Aug 2 18:15:36 1996
From: ignelzi at umich.edu (Michael A. Ignelzi, Jr.)

Subject: Transgenic Barriers
Message-ID: <v02130501ae27d9f766f8@[141.211.157.44]>

Modified barrier, access restrictions, gowning, isolators, (+/-
autoclaving), Univ of Michigan School of Dentistry (transgenic and
knockout
colonies)



>Hi folks,
>
>I am trying to get a feel for how many institutions are using barrier
>conditions
>for their transgenic/knockout colonies. Please reply to me off list
and I'll
>sumarize for the group. Don't forget to include where you're from!
>
>no/low barrier = conventional caging and access (+/- filter tops on
caging)
>
>modified barrier = access restrictions, gowning, isolators, (+/-
autoclaving)
>
>full barrier = shower in, isolator caging, autoclaved food and bedding
>
>Thanks in advance!
>Allison Treloar
>Technical Supervisor - Transgenics
>Bristol-Myers Squibb
>
>Allison_F._Treloar@ccmail.bms.com
>




From UCIVET at uci.edu Fri Aug 2 15:21:06 1996
From: UCIVET at uci.edu (Clifford R ROBERTS)

Subject: Transgenic Barriers
Message-ID: <9607028390.AA839020866@gandalf.bio.uci.edu>


low barrier: microisolator cages, changed in Class II hood, Masks, lab
coats,
gloves.

Cliff Roberts
Univ Calif, Irvine
_______________________________________________________________________
________
Subject: Transgenic Barriers
From:    transgenic-list@ic.ac.uk at biosmtp
Date:    8/2/96 1:19 PM

Hi folks,

I am trying to get a feel for how many institutions are using barrier
conditions

for their transgenic/knockout colonies. Please reply to me off list
and I'll
sumarize for the group. Don't forget to include where you're from!

no/low barrier = conventional caging and access (+/- filter tops on
caging)

modified barrier =   access restrictions, gowning, isolators, (+/-
autoclaving)

full barrier = shower in, isolator caging, autoclaved food and bedding

Thanks in advance!
Allison Treloar
Technical Supervisor - Transgenics
Bristol-Myers Squibb

Allison_F._Treloar@ccmail.bms.com
From pinkert at cmed.bhs.uab.edu Fri Aug 2 20:17:28 1996
From: pinkert at cmed.bhs.uab.edu (Dr. C.A. Pinkert)

Subject: Transgenic Barriers
Message-ID: <19960803002112601.AAD148@cap1>

> Date:           Fri, 02 Aug 96 14:21:06 PST
> To:             transgenic-list@ic.ac.uk
> Subject:        Re: Transgenic Barriers
> Reply-to:       transgenic-list@ic.ac.uk
After pulling up a number of responses to the barrier question - with
each individual describing a facility, kindly respond directly to
Allison Treloar (Allison_F._Treloar@ccmail.bms.com) as it is getting
to be a bit much.



From p.sobieszczuk at ic.ac.uk Sat Aug 3 16:53:41 1996
From: p.sobieszczuk at ic.ac.uk (Peter Sobieszczuk)

Subject: "TgList Backstage" (admin. news *1)
Message-ID: <v01510100ae28fc43f22d@[155.198.45.54]>

Dear Subscribers,

Welcome to the List, again.

The List is now about 10 days old, and quite frankly, your
administrator
(listowner) has by now much better understanding for the guys who
created
e.g. Frankenstain etc., it's easier to create the beast than to control
it.

On a positive side: The List has (as of 1st of Aug.) approximately 127
subscribers, surprisingly only, less than 40% from the old rodent-
research
list.

Congratulation to Claire and Allison for their first and second posting
(world debut on day 1 or 2 of the List existence), I will repeat their
messages below, since there were only very few people subscribed at
that
time.

On a technical note: function "reply-to" the list, was set up
deliberately
to make sending replies easier for people who are not that familiar
with
the electronics of this game, but as C.A. Pinkert noticed, this could
be
too much on occasions.

Therefore, an explanation: if you use "reply" function of your e-mail
package (e.g. "reply" from "message" menu in Eudora) remember it will
go
back to the list, not only to the original sender. To send your reply
(or
any message) just to one person use "new" message or type that address
in
the "to:" field yourself.

It would also appear to be sensible to cut out everything what is not
relevant from the copy of the original letter in your "reply to the
list"
message.

And try to remember to always fill the subject field in the new
postings as
well as signing your message (makes life easier for everyone).

Hope this will be of some help, and wishing you happy postings.


Peter
p.sobieszczuk@ic.ac.uk


P.S.

Copies of the first two posting from the last week:
____________________________________________________________________
Hello Transgenic List Subscribers:

Can anyone direct me to any studies, papers, or general information,
regarding stress indicators of mice that have been placed in the
dark or light reduced areas for 24 hours or longer ?

Thank you

Claire Smits
csmits@qlt-pdt.com
_____________________________________________________________________
I hope there are enough of you signed up on this new list to help me
out..

Aside from Genome Systems, does anyone know of other suppliers of
contract
knock-out mouse production services. I don't know if DNX has this
service up
and running yet.

Thanks,

Allison
Technical Supervisor-Transgenics
Bristol-Myers Squibb Co.
Allison_F._Treloar@ccmail.bms.com
_____________________________________________________________________




From MEISLERM at hg-basic1mail.hg.med.umich.edu Sat Aug 3 14:04:43
1996
From: MEISLERM at hg-basic1mail.hg.med.umich.edu (MEISLERM)

Subject: brain and neuron specific promoters
Message-ID: <A145386529@HG-BASIC1MAIL.HG.MED.UMICH.EDU>

Does anyone know of a recent review of promoters with in vivo
specificity for subsets
of neurons, glia, or brain regions? We are looking for promoters
specific for brain
regions, spinal cord, motor neurons, etc.
Thanks for your suggestions.



Miriam Meisler, Ph. D.
Department of Human Genetics
University of Michigan
4708 Medical Sciences II
Ann Arbor, MI 48109-0618
tel 313 763 5546; FAX 313 763 9691.email: meislerm@umich.edu



From gstuart at UVic.CA Sat Aug 3 18:01:42 1996
From: gstuart at UVic.CA (Greg Stuart)

Subject: Rat liver anatomy question
Message-ID: <2.2.32.19960804000142.006a7bb4@pop.uvic.ca>

Hello - I'd like to draw on the collective experience of the
subscribers.
Can anybody describe where the following regions of the rat liver are
located?

- centrilobular region
- midzonal hepatocytes
- periportal regions

Also, are these regions found in each lobe of the liver, or are they
specific
to particular lobes or regions?

Please respond directly, thanks. Cheers, Greg :)

--
Greg Stuart    gstuart@uvic.ca
Centre for Environmental Health .   .*#*.            .*#*.   .*#*.
Department of Biology    * | | | * | | | *        * | | | * | | | *
University of Victoria * | | | *   * | | | *    * | | | *   * | | | *
P.O. Box 3020         * | | | *        * | | | * | | | *        * | |
|*
Victoria, B.C., CANADA `*#*'            `*#*'    `*#*'           `*#*'
V8W 3N5.
Tel (604)472-4067, Fax(604)472-4075
http://darwin.ceh.uvic.ca/students/gstuart/




From hage at med.unc.edu Sat Aug 3 23:01:14 1996
From: hage at med.unc.edu (John Hagaman)

Subject: Transgenic Barriers
Message-ID: <v01540b01ae297598a9dd@[152.2.169.3]>

>Hi folks,
>
>I am trying to get a feel for how many institutions are using barrier
>conditions
>for their transgenic/knockout colonies. Please reply to me off list
and I'll
>sumarize for the group. Don't forget to include where you're from!

we us modified barrier =   access restrictions, gowning, isolators, (+/-
autoclaving, both )

>
>Allison_F._Treloar@ccmail.bms.com
>

John R. Hagaman
Pathology Department
University of North Carolina Medical School
7525 Brinkhous-Bullitt, Room 703
Chapel Hill, N.C. 27705

966-6912 office
966-8800 Fax
From kelly at citi2.fr Mon Aug   5 15:12:58 1996
From: kelly at citi2.fr (U344)

Subject: mouse testosterone levels
Message-ID: <v02120d02ae2c09f00f80@[192.70.98.85]>

        Hello, does anyone know of a way to measure mouse testosterone
levels, or of anyone who is? I'd really like to avoid having to set up
an
ELISA for a few samples...
                                cheers...

INSERM Unite 344 Endocrinologie Moleculaire
156 rue de Vaugirard, 75730 Paris cedex 15 France

'Man... I don't EVEN have an opinion...'




From browng at medicine.wustl.edu Mon Aug 5 08:53:29 1996
From: browng at medicine.wustl.edu (Gary Brown)

Subject: WWW resource
Message-ID: <Pine.GSO.3.94.960805074227.5164A-100000@medicine>

Hi,

Just a brief wee message here from St Louis. At the risk of tooting my
own
horn I'd like to let anyone interested know about my microinjection /
transgenisis related homepage. Apologies go to those who may have seen
this on the COMPMED listings, or to those who subscribe to this list
after
my listing THIS list on the page in question! Anyways, here it is...

Page Name: The Microinjection Workshop
Page address:
http://ourworld.compuserve.com/homepages/TheBroons/homepage.htm
How best to find: Although I have registered with most of the widely
used
search engines, Yahoo is still a problem.. Best to use the Starting
Point
(www.stpt.com) and select the combination Metacrawler engine searching
on
"microinjection".

I'm trying to build up an E-mail mailing list to alert when the page is
updated (approx. 1 x per month). If interested please reply OFF-LIST
tothe
following:

75017.1050@compuserve.com
Thanx :-)

Gary




From Allison_F._Treloar at ccmail.bms.com Mon Aug 5 10:59:37 1996
From: Allison_F._Treloar at ccmail.bms.com (Allison F. Treloar)

Subject: Summary: Tg Barriers
Message-ID: <9607058392.AA839265404@ccgate1.bms.com>

First and foremost, a big thanks to everyone who responded to my
question
regarding barrier conditions for trangenic colonies. Here's the
outcome:

17 respondents
    3 facilties use low/no barrier conditions
    10 facilities use modified barrier conditions
    4 facilities use full barrier conditions


low/no barrier respondents stated that these conditions were used
primarily for
single transgenic lines or areas where investigators could generally
access
transgenic stock animals.

modified barrier users seem to rely on access restrictions, gowing,
dedicated
staff, isolator caging and autoclaved food/bedding, but don't go as far
as
shower-in or sterile clothing/scrubs. Most cage changing/animal
manipulation is
done in laminar flow hoods. This is certainly the most popular,
flexible and
cost effective option based on current opinion.

full barriers were used by a few facilities and included air showers,
sterile
clothing/scub requirements as well as all the modified barrer
conditions. No
respondents with full barriers used shower-in.

Thanks, again for all your help.

Allison

Allison_F._Treloar@ccmail.bms.com
From gina_seeburger at Merck.Com Mon Aug 5 15:07:08 1996
From: gina_seeburger at Merck.Com (Gina Seeburger)

Subject: "TgList Backstage" (admi
Message-ID: <199608052001.QAA29238@igw2>

                                                  RE>"TgList Backstage"
(admin.


--------------------------------------
Date: 8/3/96 10:56 AM
To: Gina Seeburger
From: transgenic-list@ic.ac.uk
Dear Subscribers,

Welcome to the List, again.

The List is now about 10 days old, and quite frankly, your
administrator
(listowner) has by now much better understanding for the guys who
created
e.g. Frankenstain etc., it's easier to create the beast than to control
it.

On a positive side: The List has (as of 1st of Aug.) approximately 127
subscribers, surprisingly only, less than 40% from the old rodent-
research
list.

Congratulation to Claire and Allison for their first and second posting
(world debut on day 1 or 2 of the List existence), I will repeat their
messages below, since there were only very few people subscribed at
that
time.

On a technical note: function "reply-to" the list, was set up
deliberately
to make sending replies easier for people who are not that familiar
with
the electronics of this game, but as C.A. Pinkert noticed, this could
be
too much on occasions.

Therefore, an explanation: if you use "reply" function of your e-mail
package (e.g. "reply" from "message" menu in Eudora) remember it will
go
back to the list, not only to the original sender. To send your reply
(or
any message) just to one person use "new" message or type that address
in
the "to:" field yourself.

It would also appear to be sensible to cut out everything what is not
relevant from the copy of the original letter in your "reply to the
list"
message.
And try to remember to always fill the subject field in the new
postings
as
well as signing your message (makes life easier for everyone).

Hope this will be of some help, and wishing you happy postings.


Peter
p.sobieszczuk@ic.ac.uk


P.S.

Copies of the first two posting from the last week:
____________________________________________________________________
Hello Transgenic List Subscribers:

Can anyone direct me to any studies, papers, or general information,
regarding stress indicators of mice that have been placed in the
dark or light reduced areas for 24 hours or longer ?

Thank you

Claire Smits
csmits@qlt-pdt.com
_____________________________________________________________________
I hope there are enough of you signed up on this new list to help me
out..

Aside from Genome Systems, does anyone know of other suppliers of
contract
knock-out mouse production services. I don't know if DNX has this
service
up
and running yet.

Thanks,

Allison

Technical Supervisor-Transgenics
Bristol-Myers Squibb Co.
Allison_F._Treloar@ccmail.bms.com
_____________________________________________________________________
From TSAUNDER at hg-basic1mail.hg.med.umich.edu Tue Aug 6 10:57:55
1996
From: TSAUNDER at hg-basic1mail.hg.med.umich.edu (TSAUNDER)

Subject: Meeting Announcement
Message-ID: <E62DFE6902@HG-BASIC1MAIL.HG.MED.UMICH.EDU>

THE UNIVERSITY OF MICHIGAN CENTER FOR ORGANOGENESIS
1st International Symposium

Molecular Control of Organogenesis

Saturday, October 5, 1996
Horace H. Rackham Graduate School Amphitheater
The University of Michigan
Ann Arbor, Michigan


8:45-9:00      Opening Remarks, Dr. Giles G. Bole, Dean,
   The University of Michigan Medical School Dean

9:00-9:50      Dr. Eric Olson, University of Texas, Dallas, Texas USA,
               "Genetic control of myogenesis"

9:50-10:40     Dr. Peter Gruss, Max Planck Institute of Biophysical
                            Chemistry, Germany,
  "Genes required for the development of the visual system"

10:40-10:50    Break

10:50-11:40    Dr. Cliff Tabin, Harvard Medical School, Boston,
                             Massachusetts, USA,
   "Signals patterning the vertebrate limb"

11:40-12:30    Dr. Yuh Nung Jan, University of California, San
Francisco,
                              California, USA,
    "Molecular control of neural development"

12:30-2:30     Poster Session (lunch provided)

2:30-3:20      Dr. Paul Sternberg, California Institute of Technology,
                              Pasadena, California, USA,
   "Precise formation of a hole: the C. elegans vulva"

3:20-4:10      Dr. Richard M. Harland, University of California,
Berkeley,
                                  California, USA,
   "Re-use of signaling molecules in vertebrate axis formation and
                  organogenesis"

4:10-4:20         Closing Remarks, Dr. Seigo Izumo, Acting Director,
Center for
                  Organogenesis, The University of Michigan


Friday, October 4, 1996
   5:30-7:00 p.m.      Meet the Speakers at an Open Reception,
                       Museum of Art, The University of Michigan

For further information, please contact Michelle Shukait at 936-2499 or
mshukait@umich.edu




From hage at med.unc.edu Wed Aug 7 00:28:28 1996
From: hage at med.unc.edu (John Hagaman)

Subject: White noise for Mouse Embryo Yield
Message-ID: <v01540b01ae2d819d1d72@[152.2.169.1]>

Does anyone have experience re: noise in mouse rooms and how it may
affect
the yield of embryos? Do mice "get used to certain noises" Does with
drawal or introduction of noise change the yield?

Responses off list will be summarized.

Thanks

John R. Hagaman
Pathology Department
University of North Carolina Medical School
7525 Brinkhous-Bullitt, Room 703
Chapel Hill, N.C. 27705

966-6912 office
966-8800 Fax




From fmargoli at umabnet.ab.umd.edu Wed Aug 7 10:38:50 1996
From: fmargoli at umabnet.ab.umd.edu (Frank L. Margolis)

Subject: Knock out master list
Message-ID: <v01530501ae2e5aacaf10@[134.192.49.38]>

Thanks to all for your responses to my query, they were most helpful.

Frank Margolis

Frank L. Margolis Ph.D.
Department of Anatomy
University of Maryland at Baltimore
School of Medicine
685 West Baltimore Street
Baltimore MD 21201

Phone 410-706-8913 office
             -8914 lab
FAX          -2512 Department office




From r-carver at nimr.mrc.ac.uk Thu Aug 8 17:47:28 1996
From: r-carver at nimr.mrc.ac.uk (Rick Carver)

Subject: LASA Transgenics Meeting
Message-ID: <4080.9608071557@nimsn01.nimr.mrc.ac.uk>

Dear All

I am now able to give details of the speakers for the forthcoming LASA
Transgenics meeting.

Transgenesis- setting the scene               Dr   Roy Forster, Transgene
Transgenic Animals in Developmental Biology   Dr   Reter Rigby, NIMR
Transgenic Animals in Immunology              Dr   Mike Owen, ICRF
Transgenic Animals in Production Proteins     Dr   Ron James,
Pharmaceutical
Proteins Ltd
Transgenic Animals in Neuroscience            Prof Rick Lathe, Univ of
Edinburgh
Transgenic Animals in Oncology                Dr Frances Balkwell, ICRF
Transgenic Animals in Pharmaceuticals         Dr John McNeish, Pfizer
US
   Discovery and Development
Ethical and Legal Aspects of the Use of       Dr Kenneth Boyd, Univ of
Edinburgh
   Transgenic Animals


The cost will be 55 pounds stirling for LASA members and 75 pounds
stirling
for non-members. Registration forms are available from:    LASA
Speciality
Sections, PO Box 3993, Tamworth, Staffs, B78 3QU, UK. The closing date
for
registrations is 23 Aug 96.



Rick Carver
Head of Biological Services and Institute Veterinarian
National Institute for Medical Research
The Ridgeway
MILL HILL
London                     E-Mail: r-carver@nimr.mrc.ac.uk
NW7 1AA                    Tel:   + 44 (0)181 959 3666 ext 2199
United Kingdom             Fax:   + 44 (0)181 913 8601

"There is something fascinating about science: one gets such wholesale
returns of conjecture out of such a trifling investment of fact" - Mark
Twain




From gac at po.mri.montana.edu Wed Aug 7 18:36:49 1996
From: gac at po.mri.montana.edu (George Carlson)

Subject: discontinuation of strains
Message-ID: <v01540b06ae2ed353bf48@[204.200.84.36]>

Most of the intra-H-2 recombinant B10 congenic strains developed by the
late Dr. Jack Stimpfling of McLaughlin Research Institute will be
discontinued as live stock in the near future. Following Dr.
Stimpfling's
retirement, a grant from the American Cancer Society allowed us to
complete
isolation of his more recently identified recombinant chromosomes, to
refine localization of the crossover sites within H-2, to delimit the
congenic intervals for each strain, and to preserve the strains for
future
use. We are now in the final stage of this project and expect to have
all
strains frozen before January 1997. DNA samples will continue to be
available.

The strains are listed in:
Turner et al. Meiotic recombination within the H-2K--H-2D interval:
characterization of a panel of congenic mice, including 12 new strains,
using DNA markers. Immunogenetics 1993, 38:332-340.

Additional data on our congenic panel are being prepared for
publication.

Breeding pairs are available for any investigator who requests them. We
only ask that investigators who "adopt" a strain make it available to
others in the community and provide notification if they stop the line.

I will provide a listing of the strains that we will continue as live
stock
after we get a better idea of interest. Please let me know if you need
any
of these mice. Suggestions of other appropriate lists for posting this
notice would also be appreciated.

Thank you.

George A. Carlson (gac@po.mri.montana.edu)
McLaughlin Research Institute
1520 23rd Street South
Great Falls, Montana 59405
406 452 6208, fax: 454 6019 or 454 6004
From S.Tan at Anatomy.Unimelb.EDU.AU Thu Aug 8 20:41:53 1996
From: S.Tan at Anatomy.Unimelb.EDU.AU (Seong Seng Tan)

Subject: White noise for Mouse Embryo Yield
Message-ID: <v01540b04ae2f58725c88@[128.250.225.71]>

>Does anyone have experience re: noise in mouse rooms and how it may
affect
>the yield of embryos? Do mice "get used to certain noises" Does
with
>drawal or introduction of noise change the yield?
>

  We are currently undergoing major building renovations and this week
alone, three of our litters have been eaten by their mothers. We
suspect
loud sudden noises are responsible.

Seong-Seng Tan

=======================================================================
===
Senior Lecturer, Department of Anatomy & Cell Biology
The University of Melbourne
Parkville 3052
Victoria, Australia
Fax 61 3 9 347 5219, Phone 61 3 9 344 5787
Web: http://www.anatomy.unimelb.edu.au

=======================================================================
===




From fmargoli at umabnet.ab.umd.edu Thu Aug 8 11:11:34 1996
From: fmargoli at umabnet.ab.umd.edu (Frank L. Margolis)

Subject: Lists
Message-ID: <v01530502ae2faf1c9718@[134.192.49.38]>

To all-

In response to several requests I am posting a summary of all the info
I
have received about the availability of lists of Knockouts and
transgenics
and specific strains (without any editorial evaluation).

BioMedNet   has at least a partial list of knockouts
JAX homepage is http://www.jax.org

Current list of induced mutant strains accepted for inclusion in the
Induced Mutant Resource (IMR) at JAX is on the IMR home page-
http://lena.jax.org/resources/documents/imr/

An index of strains is available at-
http://lena.jax.org/resources/documents/imr/imr.html

and more details at-
http://lena.jax.org/resources/documents/imr/imrdata.html

for queries about any JAX mice contact-
micetech@jax.org


Also the following web sites have info that may prove to be useful-
NIH transgenic directory   Tbase
http://www.gdb.org/Dan/tbase/tbase.html

Mouse and rat research home page-
http://www.cco.caltech.edu/~mercer/htmls/rodent_page.html#homepages

and as was also pointed out to me: 1-a Medline search for info on
specific
genes or strains as well as 2- right here where I started all this.


Thanks again for all your input.

Frank Margolis

Frank L. Margolis Ph.D.
Department of Anatomy
University of Maryland at Baltimore
School of Medicine
685 West Baltimore Street
Baltimore MD 21201

Phone 410-706-8913 office
             -8914 lab
FAX          -2512 Department office




From gstuart at UVic.CA Thu Aug 8 08:37:43 1996
From: gstuart at UVic.CA (Greg Stuart)

Subject: Isolation of bone marrow & colon tissues
Message-ID: <2.2.32.19960808143743.006bd2ac@pop.uvic.ca>

Hello everyone. Does anybody have protocols that they could share for
the
isolation of rat (or rodent) bone marrow and colon tissue? Thanks, Greg
:)
--
Greg Stuart    gstuart@uvic.ca
Centre for Environmental Health .   .*#*.            .*#*.   .*#*.
Department of Biology    * | | | * | | | *        * | | | * | | | *
University of Victoria * | | | *   * | | | *    * | | | *   * | | | *
P.O. Box 3020         * | | | *        * | | | * | | | *        * | |
|*
Victoria, B.C., CANADA `*#*'            `*#*'    `*#*'           `*#*'
V8W 3N5.
Tel (604)472-4067, Fax(604)472-4075
http://darwin.ceh.uvic.ca/students/gstuart/




From r-lovell at nimr.mrc.ac.uk Fri Aug 9 18:38:45 1996
From: r-lovell at nimr.mrc.ac.uk (Robin Lovell-Badge)

Subject: Copy of: Re: White noise for Mouse Embryo Yield
Message-ID: <v01530500ae3117938cc0@[192.107.168.62]>

Dear All

There is considerable published and anecdotal stuff on noise in animal
rooms. Excessive noise, e.g. from building works, can have all sorts
of
effects which will lower reproductive efficiency. Mice seem to
particularly dislike sudden noises, so it can be beneficial to have
music
playing (classical or modern, but not too much rap !) during the day
when
cages are being changed, etc. But again, it should not be very loud,
just
enough to mask the dropping of cages, screams of students, etc.

If anyone is desperate I can try to locate published work on this - but
it
is not recent and may take a while.

Robin Lovell-badge




>You wrote:
>
>>Does anyone have experience re: noise in mouse rooms and how it may
affect
>>the yield of embryos? Do mice "get used to certain noises" Does
with
>>drawal or introduction of noise change the yield?
>>
>
>       While working at the University of Kentucky, I found a number
of
>interuptions in breeding colonies due to the noise caused by
construction.
>Liters were either canabalized by the mother or breeding stopped
>completely. Hope this helps.
>
>
>----------------------------------------------------------------------
---
>                                             ( )_( )
> Richard S. Cluck, B.S., LATG                   o o
> Supervisor, VMU, VA, Lexington               ==\o/==
> Research 151                                    /\    )
> Cooper Drive                                   / \ (
> Lexington, KY 40511                           /    \)
> Phone: 606-281-4927
> Fax: 606-281-4989
> E-Mail: cluck,richard_@lexington.va.gov
>----------------------------------------------------------------------
---




From cfoltz at MIT.EDU Mon Aug 12 12:50:40 1996
From: cfoltz at MIT.EDU (Charmaine Foltz )

Subject: control of P. pneumotropica
Message-ID: <9608121550.AA28963@MIT.MIT.EDU>

I am currently treating a small colony of Rag-2 mice to try to
eliminate
Pasteurella pneumotropica, which has been a significant pathogen for
this
particular genotype. My question to the group is how many of you have
had
experience with doing this (I am using enrofloxicin in the water).
Additionally, having read recently that humans can colonize with the
bacteria (Cudrado-Gomez LM, et al. Pasteurella pneumotropica pneumonia
in
a patient with AIDS. Clinical Inf. Dis, 1995;21:445-6), I wonder if
anyone
on the list has had experience with a barrier facility becoming
colonized
with the agent, presumably from contamination by a human source.

Thank you,
Charmaine Foltz, DVM, Dipl. ACLAM
MIT Div. Comparative Medicine
77 Massachusetts Ave. Bldg. 45
Cambridge, MA 02139
(617)252-1804
(617)258-5708 (fax)
cfoltz@mit.edu
From dknudsen at scruznet.com Mon Aug 12 21:32:45 1996
From: dknudsen at scruznet.com (Dave Knudsen)

Subject: control of P. pneumotropica
Message-ID: <v01540b01ae35a62ecf39@[165.227.113.91]>

>I am currently treating a small colony of Rag-2 mice to try to
eliminate
>Pasteurella pneumotropica, which has been a significant pathogen for
this
>particular genotype. My question to the group is how many of you have
had
>experience with doing this (I am using enrofloxicin in the water).
>Additionally, having read recently that humans can colonize with the
>bacteria (Cudrado-Gomez LM, et al. Pasteurella pneumotropica pneumonia
in
>a patient with AIDS. Clinical Inf. Dis, 1995;21:445-6), I wonder if
anyone
>on the list has had experience with a barrier facility becoming
colonized
>with the agent, presumably from contamination by a human source.

Charmaine -

I've had some experience with this agent. In a colony of over 12,000
SCID
mice used in transplantation studies (and therefore routinely
irradiated),
we were highly motivated to remove pasteurellosis from the list. Using
draconian barrier procedures and caesarian rederivation of every strain
to
be introduced into the facility, we were able to rid all of our
colonies of
P pneumo as well as everything else I know how to test for.
Individually
ventillated caging, a frequent health monitoring system, and restricted
access help kept it that way. I'll be happy to give you more details
off
line should you be interested in this approach.

Dave Knudsen DVM, MS, DACLAM
Scotts Valley, California, USA
<dknudsen@scruznet.com>

***
"In my judgement it is either an enigma or some kind of bug. If it
dies, I
will take it apart and see what its arrangements are. I never had a
thing
perplex me so." Mark Twain, 1893 (from "Extracts from Adam's Diary")
***
From dbowtell at petermac.unimelb.edu.au Tue Aug 13 21:49:47 1996
From: dbowtell at petermac.unimelb.edu.au (David Bowtell)

Subject: Double KO with increased G418
Message-ID: <v01530501ae3699bc24e4@[203.4.164.101]>

We have used increased G418 concentration as a means of selecting
homozgous
cells on two occassions. One construct had a pGKNeo casssette and the
cells
were still laughing in 4mg/ml (we blinked first). The second construct
was
promoterless, the cells were marginally G418 resistant and it worked
like a
dream. The paper by Mortensen (MCB 12, 2391) used a pGKNeo cassette
but I
am unclear whether it was the point mutant (low activity) neo cassette.
So
we would be interested in other people's experience with this approach
to
double KO in ES cells (and any tidbits on experience targeting the
second
allele indepndantly).

Many thanks.

David Bowtell.

******* Please note new email address **********
David Bowtell
Principal Research Fellow
Research Division
Peter MacCallum Cancer Institute
Locked Bag 1 A'Beckett St,
Melbourne 3000

Lab 61-3-96561287
Office 61-3-96561296
Fax 61-3-96561411
email: dbowtell@petermac.unimelb.edu.au




From wixson at biovax.dnet.basf-ag.de Tue Aug 13 20:16:55 1996
From: wixson at biovax.dnet.basf-ag.de (Sally Wixson, VMD, MS)

Subject: Knockout advice
Message-ID: <9608131716.AA23717@gwr.zxd.basf-ag.de>

    I direct the Lab Animal department of a pharmaceutical research
company
that has an active, but disorganized, knock out team. I have been here
eight
months and trying to get their animal work back on track. I would be
grateful
for your advice on several topics:
.....what strain of vactomized males do you use and why? How do you
assess
their sterility (other than ordering then vasec. by the vendor)? How
often do
you use vasec. males? (my invesitagors use each male once/week or
less!)

.....what strain of embryo recipients do you use for ES cell work? Our
donors
are C57BL/6, recipients are B6D2F1's. My team wants a "big" target so
they keep
the females here three months or so on a high fat diet so they are big
enough
to inject. Obviously, this leads to a waste of cage space. Many people
use
ICR's or CD1's as recipients because they are a big target and good
moms!

.....in breeding chimeras, our PI insists that each chimera must
produce100
pups (black) before being considered a failure at germ line
transmission. Many
of the chimeras are lowly sterile or sterile, so this format entails
breeding
formany months to more than a year. Several other labs told me their
chimeras
get three matings and if no agouti pups are produced, they are ENA'd.

.....do you produce your own ES cells or get them from
donation/purchase from
other labs? We are currently buying the RW4 ES cell line from Genome
Systems.
Do you require MAP testing of the new ES cell lines?

.....do you superovulate your females for embryo donation? My PI says
this doe
not yield viable embryos for ES cell work, so they breed 4-5X the
number of
females they rpedict will be needed for each injectionso that they have
enough
embryos.

Many thanks for the advice,
Sally K, Wixson, VMD, MS



From wixson at biovax.dnet.basf-ag.de Tue Aug 13 20:16:55 1996
From: wixson at biovax.dnet.basf-ag.de (Sally Wixson, VMD, MS)

Subject: Knockout advice
Message-ID: <9608131716.AA23717@gwr.zxd.basf-ag.de>

    I direct the Lab Animal department of a pharmaceutical research
company
that has an active, but disorganized, knock out team. I have been here
eight
months and trying to get their animal work back on track. I would be
grateful
for your advice on several topics:
.....what strain of vactomized males do you use and why? How do you
assess
their sterility (other than ordering then vasec. by the vendor)? How
often do
you use vasec. males? (my invesitagors use each male once/week or
less!)

.....what strain of embryo recipients do you use for ES cell work? Our
donors
are C57BL/6, recipients are B6D2F1's. My team wants a "big" target so
they keep
the females here three months or so on a high fat diet so they are big
enough
to inject. Obviously, this leads to a waste of cage space. Many people
use
ICR's or CD1's as recipients because they are a big target and good
moms!

.....in breeding chimeras, our PI insists that each chimera must
produce100
pups (black) before being considered a failure at germ line
transmission. Many
of the chimeras are lowly sterile or sterile, so this format entails
breeding
formany months to more than a year. Several other labs told me their
chimeras
get three matings and if no agouti pups are produced, they are ENA'd.

.....do you produce your own ES cells or get them from
donation/purchase from
other labs? We are currently buying the RW4 ES cell line from Genome
Systems.
Do you require MAP testing of the new ES cell lines?

.....do you superovulate your females for embryo donation? My PI says
this doe
not yield viable embryos for ES cell work, so they breed 4-5X the
number of
females they rpedict will be needed for each injectionso that they have
enough
embryos.

Many thanks for the advice,
Sally K, Wixson, VMD, MS



From dbowtell at petermac.unimelb.edu.au Tue Aug 13 21:49:47 1996
From: dbowtell at petermac.unimelb.edu.au (David Bowtell)

Subject: Double KO with increased G418
Message-ID: <v01530501ae3699bc24e4@[203.4.164.101]>
We have used increased G418 concentration as a means of selecting
homozgous
cells on two occassions. One construct had a pGKNeo casssette and the
cells
were still laughing in 4mg/ml (we blinked first). The second construct
was
promoterless, the cells were marginally G418 resistant and it worked
like a
dream. The paper by Mortensen (MCB 12, 2391) used a pGKNeo cassette
but I
am unclear whether it was the point mutant (low activity) neo cassette.
So
we would be interested in other people's experience with this approach
to
double KO in ES cells (and any tidbits on experience targeting the
second
allele indepndantly).

Many thanks.

David Bowtell.

******* Please note new email address **********
David Bowtell
Principal Research Fellow
Research Division
Peter MacCallum Cancer Institute
Locked Bag 1 A'Beckett St,
Melbourne 3000

Lab 61-3-96561287
Office 61-3-96561296
Fax 61-3-96561411
email: dbowtell@petermac.unimelb.edu.au




From dknudsen at scruznet.com Mon Aug 12 21:32:45 1996
From: dknudsen at scruznet.com (Dave Knudsen)

Subject: control of P. pneumotropica
Message-ID: <v01540b01ae35a62ecf39@[165.227.113.91]>

>I am currently treating a small colony of Rag-2 mice to try to
eliminate
>Pasteurella pneumotropica, which has been a significant pathogen for
this
>particular genotype. My question to the group is how many of you have
had
>experience with doing this (I am using enrofloxicin in the water).
>Additionally, having read recently that humans can colonize with the
>bacteria (Cudrado-Gomez LM, et al. Pasteurella pneumotropica pneumonia
in
>a patient with AIDS. Clinical Inf. Dis, 1995;21:445-6), I wonder if
anyone
>on the list has had experience with a barrier facility becoming
colonized
>with the agent, presumably from contamination by a human source.

Charmaine -

I've had some experience with this agent. In a colony of over 12,000
SCID
mice used in transplantation studies (and therefore routinely
irradiated),
we were highly motivated to remove pasteurellosis from the list. Using
draconian barrier procedures and caesarian rederivation of every strain
to
be introduced into the facility, we were able to rid all of our
colonies of
P pneumo as well as everything else I know how to test for.
Individually
ventillated caging, a frequent health monitoring system, and restricted
access help kept it that way. I'll be happy to give you more details
off
line should you be interested in this approach.

Dave Knudsen DVM, MS, DACLAM
Scotts Valley, California, USA
<dknudsen@scruznet.com>

***
"In my judgement it is either an enigma or some kind of bug. If it
dies, I
will take it apart and see what its arrangements are. I never had a
thing
perplex me so." Mark Twain, 1893 (from "Extracts from Adam's Diary")
***




From cfoltz at MIT.EDU Mon Aug 12 12:50:40 1996
From: cfoltz at MIT.EDU (Charmaine Foltz )

Subject: control of P. pneumotropica
Message-ID: <9608121550.AA28963@MIT.MIT.EDU>

I am currently treating a small colony of Rag-2 mice to try to
eliminate
Pasteurella pneumotropica, which has been a significant pathogen for
this
particular genotype. My question to the group is how many of you have
had
experience with doing this (I am using enrofloxicin in the water).
Additionally, having read recently that humans can colonize with the
bacteria (Cudrado-Gomez LM, et al. Pasteurella pneumotropica pneumonia
in
a patient with AIDS. Clinical Inf. Dis, 1995;21:445-6), I wonder if
anyone
on the list has had experience with a barrier facility becoming
colonized
with the agent, presumably from contamination by a human source.

Thank you,
Charmaine Foltz, DVM, Dipl. ACLAM
MIT Div. Comparative Medicine
77 Massachusetts Ave. Bldg. 45
Cambridge, MA 02139
(617)252-1804
(617)258-5708 (fax)
cfoltz@mit.edu




From browng at medicine.wustl.edu Wed Aug 14 08:52:59 1996
From: browng at medicine.wustl.edu (Gary Brown)

Subject: Knockout advice
In-Reply-To: <9608131716.AA23717@gwr.zxd.basf-ag.de>
Message-ID: <Pine.GSO.3.94.960814071607.1135A-100000@medicine>

Hi,

Just throwing in my 2 cents worth...

On Tue, 13 Aug 1996, Sally Wixson, VMD, MS wrote:

> .....what strain of vactomized males do you use and why? How do you
assess
> their sterility (other than ordering then vasec. by the vendor)? How
often do
> you use vasec. males? (my invesitagors use each male once/week or
less!)

I use Swiss Webster VAS-X males, ordered in from Taconic Farms. In my
previous position we did our own vasectomies on C57Bl/6J males
(pronuclear
microinjection only) and assessed their sterility by co-housing with
Swiss
Webster (or other inexpensive white mouse strain) females (2) for
approx.
6 weeks prior to use. Males were vasectomized at 4-5 weeks of age,
excising a minimum of 1cm of vas deferens per side.
 >
> .....what strain of embryo recipients do you use for ES cell work?
Our donors
> are C57BL/6, recipients are B6D2F1's. My team wants a "big" target so
they keep
> the females here three months or so on a high fat diet so they are
big enough
> to inject. Obviously, this leads to a waste of cage space. Many
people use
> ICR's or CD1's as recipients because they are a big target and good
moms!

I have used B6D2F1s for recipients although now use Swiss Webster
(occasionally ICR) females as they are a relatively large mouse strain.
As for feeding them on a high fat diet I personally do not subscribe to
the 'big target' philosophy - at least no bigger than using regular
mouse
chow for my chosen strain. Since we have a pool of SW females from
which
to pick those in estrus and individuals may be passed over several
times,
high fat diet pushes some of them to a size where surgery becomes more
difficult due to the increased fat deposits around the oviduct/ovary/UT
junction, and I believe that the increased incision size in the body
wall
that this necessitates increases the risk to the animal.
 >
> .....in breeding chimeras, our PI insists that each chimera must
produce100
> pups (black) before being considered a failure at germ line
transmission. Many
> of the chimeras are lowly sterile or sterile, so this format entails
breeding
> formany months to more than a year. Several other labs told me their
chimeras
> get three matings and if no agouti pups are produced, they are ENA'd.

This is insane! My initial reaction would be to tell your PI to get a
life! Our policy is to discard all female chimeras (low germline
transmission, increased sterility in the F1 generation when successful)
and to discard all >50% (by color coat) male chimeras. 3 matings
(strikes) and you're out sounds appropriate to me.
 >
> .....do you produce your own ES cells or get them from
donation/purchase from
> other labs? We are currently buying the RW4 ES cell line from Genome
Systems.
> Do you require MAP testing of the new ES cell lines?

Our lab has approached other labs for ES cells, although we are trying
to
establish our own cell line currently. With reference to the RW4 cell
line, I have used this effectively on several occasions and note that
it
has a high color coat conversion rate. I also know the individual who
created this cell line, and can forward any questions you may have to
her.
Our institution requires MAP testing of all ES cell lines for use in
mice.
>
> .....do you superovulate your females for embryo donation? My PI says
this doe
> not yield viable embryos for ES cell work, so they breed 4-5X the
number of
> females they rpedict will be needed for each injectionso that they
have enough
> embryos.
>
Yes, I superovulate females for embryo donation. I typically
superovulate
7 C57Bl/6J females for one day's effort, with a variation between 20-25
morula per plugged female. I culture these in the incubator overnight
to
blastocysts with an 85-95% conversion rate. The females used are
between
3.5-4.5 weeks old. I use Sigma Chemical Co. new embryo tested PMSG and
HCG. My stud males are mated no more than once weekly. There is a
reference for superovulating females for blastocyst production, but I
would have to look it up on Medline. Sounds to me like your PI needs
to
do some homework!

Hope this helps!


Gary Brown
E-mail: 75017.1050@compuserve.com or browng@medicine.wustl.edu
Web: http://ourworld.compuserve.com/homepages/TheBroons/
                "The Microinjection Workshop"




From kveen at nki.nl Thu Aug 15 12:52:09 1996
From: kveen at nki.nl (Karin van Veen)

Subject: blastocyst culture
Message-ID: <Pine.SOL.3.91.960815113939.13618B-100000@Hermes.nki.nl>


Hi, you all!

I hope you can help me. At our institute we do zygote injections and
blastocyst injections.
For some time now we have some problems with the blastocyst yields. It
is
either a superovulation problem or a flushing problem (or both).
Can someone give me some tips concerning the time schedule on the
superovulation. In what time intervals do you inject the hormones, and
isolate the embryo's? We tried out a lot, but can not improve our
yield.
We use C57/bl6 mice as donors.
We are also thinking, when it should be a flushing problem, to isolate
the
zygote on day 0,5, and culture them into blastocyst to be injected. Has
anyone experience on this? How long do you have to culture the zygotes
to get
blastocysts, are they good to be injected with ES cells?
All tips are welcome!

Karin.
The Netherlands Cancer Institute
Amsterdam
From browng at medicine.wustl.edu Thu Aug 15 08:23:30 1996
From: browng at medicine.wustl.edu (Gary Brown)

Subject: blastocyst culture
In-Reply-To: <Pine.SOL.3.91.960815113939.13618B-100000@Hermes.nki.nl>
Message-ID: <Pine.GSO.3.94.960815070704.14243A-100000@medicine>



On Thu, 15 Aug 1996, Karin van Veen wrote:
> Can someone give me some tips concerning the time schedule on the
> superovulation. In what time intervals do you inject the hormones,
and
> isolate the embryo's?
> We use C57/bl6 mice as donors.

We use a 12hr/12hr light-dark cycle from 4am to 4pm in our facility,
coupled with 47hrs between PMSG and HCG. These hormones are typically
given at 10.30am and (47hrs later) 9.30am repectively for this light
cycle. We use hormones obtained through Sigma Chemical Co. (Embryo
culture tested - new product) and have been obtaining yields of MORULAE
at
a rate of 20-25 per plugged C57Bl/6J female. These morulae are
cultured
overnight to blastocysts for ES cell injection in Gibco's modified
BMOC-3
culture media with pen/strep.added in a 37deg C 5% CO2 incubator.

> We are also thinking, when it should be a flushing problem, to
isolate the
> zygote on day 0,5, and culture them into blastocyst to be injected.
Has
> anyone experience on this? How long do you have to culture the
zygotes to get
> blastocysts, are they good to be injected with ES cells?
> All tips are welcome!
>
For culture past day 0.5, you would need 3 more days and some
consistently
good media. I suggest you flush morulae! Although I doubt that
flushing
would present too much of a problem, this is how I do it. Using a
short
bevel stainless 30g hypo., introduce the needle at the last oviduct
loop
before the UT junction and flush media through this area expelling the
embryos out of the uterus (cut 1cm down from the oviduct). If this is
problematical, flush through the infundibulum (requires a little more
finesse).

Hope this helps out!

Gary Brown
Email: browng@medicine.wustl.edu or 75017.1050@compuserve.com
Web: http://ourworld.compuserve.com/homepages/TheBroons/
              "The Microinjection Workshop"




From m.hampson at ic.ac.uk Thu Aug 15 18:18:40 1996
From: m.hampson at ic.ac.uk (m.hampson@ic.ac.uk)

Subject: Test - please ignore
Message-ID: <E0ur584-0001l4-00@sphinx>

Test - please ignore

--
     +-------------------------------------------------------------------
-+
     | Martyn Hampson          |    Tel:    0171 594 6973
|
     | Imperial College        |    Fax:    0171 594 6958
|
     | Computer Centre         |    E-Mail: M.Hampson@ic.ac.uk
|
     | London SW7 2BP, ENGLAND |    "Don't just do something, sit there!"
|
     +-------------------------------------------------------------------
-+




From m.hampson at ic.ac.uk Thu Aug 15 18:26:16 1996
From: m.hampson at ic.ac.uk (m.hampson@ic.ac.uk)

Subject: Test please ignore
Message-ID: <E0ur5FQ-0001lc-00@sphinx>

Test please ignore
--
   +-------------------------------------------------------------------
-+
   | Martyn Hampson          |    Tel:    0171 594 6973
|
   | Imperial College        |    Fax:    0171 594 6958
|
   | Computer Centre         |    E-Mail: M.Hampson@ic.ac.uk
|
   | London SW7 2BP, ENGLAND |    "Don't just do something, sit there!"
|
   +-------------------------------------------------------------------
-+




From m.hampson at ic.ac.uk    Thu Aug 15 18:29:17 1996
From: m.hampson at ic.ac.uk (m.hampson@ic.ac.uk)

Subject: Test - please ignore
Message-ID: <E0ur5IL-0001lj-00@sphinx>



  Test - please ignore
--
    +-------------------------------------------------------------------
-+
    | Martyn Hampson          |    Tel:    0171 594 6973
|
    | Imperial College        |    Fax:    0171 594 6958
|
    | Computer Centre         |    E-Mail: M.Hampson@ic.ac.uk
|
    | London SW7 2BP, ENGLAND |    "Don't just do something, sit there!"
|
    +-------------------------------------------------------------------
-+




From m.hampson at ic.ac.uk Thu Aug 15 18:31:25 1996
From: m.hampson at ic.ac.uk (m.hampson@ic.ac.uk)

Subject: Test - please ignore
Message-ID: <E0ur5KP-0001lr-00@sphinx>

Test - please ignore
--
   +-------------------------------------------------------------------
-+
   | Martyn Hampson          |    Tel:    0171 594 6973
|
   | Imperial College        |    Fax:    0171 594 6958
|
   | Computer Centre         |    E-Mail: M.Hampson@ic.ac.uk
|
   | London SW7 2BP, ENGLAND |    "Don't just do something, sit there!"
|
   +-------------------------------------------------------------------
-+




From gina_seeburger at Merck.Com Thu Aug 15 16:15:46 1996
From: gina_seeburger at Merck.Com (Gina Seeburger)

Subject: blastocyst culture
Message-ID: <199608160406.AAA22680@igw2.merck.com>

                                    REPLY o REPLY o REPLY o REPLY
                                    RE>blastocyst culture
Karin,
We were having problems too here in the US, but giving the first
injection(PMSG, Sigma brand) between 3:00pm and 4 (.3cc, .7.5IU) and
the
second (hCG) around 1:00 seems to help. We are also using C57BL/6
donors
for blast injections.    Collected around 7:30am. Also summer seems to
create a low.
Good luck
Gina Seeburger
Merck & Co,Inc.
gina_seeburger@merck.com

--------------------------------------
Date: 8/15/96 5:57 AM
To: Gina Seeburger
From: transgenic-list@ic.ac.uk

Hi, you all!

I hope you can help me. At our institute we do zygote injections and
blastocyst injections.
For some time now we have some problems with the blastocyst yields. It
is
either a superovulation problem or a flushing problem (or both).
Can someone give me some tips concerning the time schedule on the
superovulation. In what time intervals do you inject the hormones, and
isolate the embryo's? We tried out a lot, but can not improve our
yield.
We use C57/bl6 mice as donors.
We are also thinking, when it should be a flushing problem, to isolate
the

zygote on day 0,5, and culture them into blastocyst to be injected. Has
anyone experience on this? How long do you have to culture the zygotes
to
get
blastocysts, are they good to be injected with ES cells?
All tips are welcome!

Karin.
The Netherlands Cancer Institute
Amsterdam




From tjf at uci.edu Fri Aug 16 13:11:55 1996
From: tjf at uci.edu (Tom Fielder)

Subject: oocyte injection
Message-ID: <ae3a74350a021004d502@[128.200.21.145]>
        I am brand new to the field of transgenics, and I am setting up
a
transgenic core facility at UC-Irvine. I am still trying to optimize
my
pipets for injection of DNA into oocytes. I'm using a Sutter P-97
puller,
and 1mm OD x 0.78mm ID capillaries with internal filament. I got some
pipets that seemed to work very well yesterday, judging by the swelling
of
the pronucleus and no visible cell contents leaking out after
withdrawing
the pipet, but this morning 65% of the injected cells had died
(cytoplasm
was condensed and dark brown), while none of the uninjected controls
looked
like this. I'm afraid that the taper of the needle is too abrupt near
the
tip (i.e., it gets too big too fast). A very patient person at Sutter
is
helping me with the pipets, but my question for the list is this: Is
the
condensed, dark brown cytoplasm indicative of mechanical trauma, or is
it a
result of bad DNA/buffer, or something else?
        A second question: The books recommend using females that are
24-25 days old for superovulation. What happens if you use older ones?
Will the yield of oocytes be significantly less? I've been using 6-8
week
old FVB mice and getting about 30 eggs/mouse, but I've noticed that a
large
number of them aren't fertilized, even from the females that were
obviously
plugged. Any thoughts?
Tom

Thomas J. Fielder
UC-Irvine
University Lab Animal Resources
Transgenic Mouse Facility
147 BSA
Irvine, CA 92697-1310
e-mail: tjf@uci.edu
phone: 714-824-8579
FAX: 714-824-2003




From MKENNEDY at chmeds.ac.nz Sat Aug 17 23:12:42 1996
From: MKENNEDY at chmeds.ac.nz (MKENNEDY@chmeds.ac.nz)

Subject: Double KO with increased G418
Message-ID: <960817221242.c621@chmeds.ac.nz>

Date sent:   17-AUG-1996 22:06:32
Hi Dave,

Just got back from this year's Queenstown Meeting, which (for my
sins) I organised.

>We have used increased G418 concentration as a means of selecting
homozgous
>cells on two occassions. One construct had a pGKNeo casssette and the
cells
>were still laughing in 4mg/ml (we blinked first). The second
construct was
>promoterless, the cells were marginally G418 resistant and it worked
like a
>dream. The paper by Mortensen (MCB 12, 2391) used a pGKNeo cassette
but I
>am unclear whether it was the point mutant (low activity) neo
cassette. So
>we would be interested in other people's experience with this approach
to
>double KO in ES cells (and any tidbits on experience targeting the
second
>allele indepndantly).

For what it is worth, I sequenced Mortensen's pNTK construct, which
I am pretty sure was the one he used in the MCB paper, and it was
of the mutant neo variety. I am at home at present, but if you
haven't seen it, then John Sedivy had a very interesting MCB a
year or so back on titrating G418 to obtain high efficiency KO's,
with some interesting observations that related to the homozygous
KO method. Let me know if you need more info, and I'll dig it
out when I am at work. I have an oligo from close to the
polymorphic site in the neo gene, if you want to check other neo
constructs by sequencing - you're welcome to some of it.

Cheers,

Martin

NNNN   NN    Martin A Kennedy (E-mail = mkennedy@chmeds.ac.nz)   ZZZZZZZ
NN NN NN          Cytogenetic and Molecular Oncology Unit           ZZZ
NN NN NN              Christchurch School of Medicine             ZZZ
NN   NNNN                Christchurch, New Zealand               ZZZZZZZ
                 Phone (64-3)364-0880   Fax (64-3)364-0750



From stewarv at cesmtp.ccf.org Mon Aug 19 12:40:44 1996
From: stewarv at cesmtp.ccf.org (Valerie Stewart M.S.)

Subject: Knockout advice -Reply
Message-ID: <s2185375.030@cesmtp.ccf.org>

Hi Sally--

We use Swiss Webster vasectomized males, bought from Taconic. We
don't routinely test them, and have so far (knock on wood!) never had
a leak. When I used to do the vasectomies myself, we tested them with
two females each, let them plug both, then waited 3 weeks for
hopefully no resultant pregnancies. We use vasectomized males twice
a week to every other day. They can be used every night for a couple
of weeks before needing some down time. Once a week is ridiculous,
they don't need to keep their sperm count up after all! We use them
for a year as well, or until they stop performing. We use Swiss
Webster or CD-1 females for recipients--they are easy to estrus
select, good mothers, and cheap. I wouldn't keep them on a high fat
diet--in fact, we routinely cull out recipients that are too fat and
discard them.

As for testing germline transmission, 100 pups to "prove" lack of
transmission is the standard, but practicality usually dicatates
compromise. We only breed males over 50% chimeric, females only if
there are no male chimeras available. Three litters is a good
number, if they're good-sized litters, I usually try for 25-30 pups.

We are using E14.1 cells from Cincinnati, but I've used CCE, D3, J1,
R1, etc. Cell line is crucial; I've heard Genome Systems' line works
fine.

We superovulate for blastocysts routinely, using C57Bl6 females. We
generally do 10 at a time, with the standard amounts/timing of
hormones, and a 6:30am/6:30pm light cycle. The one thing we find is
crucial is to match body weight of the donors, 13-15g at time of PMS
administration. We do three injection days per clone, averaging 30
blastocysts per 10 donors (with the occasional null day). I have
gotten good results with number and % chimerism, as well as germline
transmission with these blastocysts. We only use the males once per
week, and usually only for 6-8 months, though a colleague says she
uses them twice a week for a year with no trouble.

Good Luck--
Valerie Stewart
Transgenic/Knockout Facility
Cleveland Clinic Research Institute
"stewarv@cesmtp.ccf.org"

>>> Sally Wixson, VMD, MS <wixson@biovax.dnet.basf-ag.de> - 8/13/96
1:16 PM >>>
    I direct the Lab Animal department of a pharmaceutical research
company that has an active, but disorganized, knock out team. I have
been here eight months and trying to get their animal work back on
track. I would be grateful for your advice on several topics:
.....what strain of vactomized males do you use and why? How do you
assess their sterility (other than ordering then vasec. by the
vendor)? How often do you use vasec. males? (my invesitagors use
each male once/week or less!)

.....what strain of embryo recipients do you use for ES cell work?
Our donors are C57BL/6, recipients are B6D2F1's. My team wants a
"big" target so they keep the females here three months or so on a
high fat diet so they are big enough to inject. Obviously, this
leads to a waste of cage space. Many people use ICR's or CD1's as
recipients because they are a big target and good moms!

.....in breeding chimeras, our PI insists that each chimera must
produce100 pups (black) before being considered a failure at germ
line transmission. Many of the chimeras are lowly sterile or
sterile, so this format entails breeding formany months to more than
a year. Several other labs told me their chimeras get three matings
and if no agouti pups are produced, they are ENA'd.

.....do you produce your own ES cells or get them from
donation/purchase from other labs? We are currently buying the RW4
ES cell line from Genome Systems. Do you require MAP testing of the
new ES cell lines?

.....do you superovulate your females for embryo donation? My PI says
this doe not yield viable embryos for ES cell work, so they breed
4-5X the number of females they rpedict will be needed for each
injectionso that they have enough embryos.

Many thanks for the advice,
Sally K, Wixson, VMD, MS




From crussell at ResGen.COM Mon Aug 19 12:03:24 1996
From: crussell at ResGen.COM (Chris Russell)

Subject: employment opportunity
Message-ID: <199608191556.KAA21398@twister.resgen.com>

Position Available Immediately!
Lab Animal Technologist/Research Assistant

Skills Required:
--Knowledgeable in theoretical embryology
--Skilled in applied embryology and microsurgery techniques
--Primarily laboratory work using the mouse as a model system
--Molecular biology experience preferred

--Salary: $28-30K plus great   benefits

--Excellent work environment in a growing biotech firm in Huntsville,
AL

Please send resume and references to:
Dr. Chris Russell
Research Genetics, Inc.
2700 Memorial Parkway, SW
Huntsville, AL 35801
1-800-711-2089
e-mail: crussell@resgen.com
***********************************************************************
****
                            Christopher G. Russell, Ph.D.
                            RESEARCH GENETICS, INC
                                Resources for Research
-_+            _+_+            ++-_        _+-_        _+_          _++
_+-_
       -+      -+     -+        +-    +-     -+   +      -+   -+           -+   +
-+
        -+     -+     -+       +-     -+    -+    -+    -+     -+         -+
-+       -+
           -++-+       -+_-+   -+_+        -++-         -+_++      -+_+
                        2700 Memorial Parkway SW
                              Huntsville, AL 35801
 Phone 1-800-533-4363, ext 2211 or Direct 1-800-711-2089
                               U.K. 0-800-89-1393
              Fax 1-205-551-1021 or 1-205-536-9016
                              crussell@resgen.com
***********************************************************************
******




From stewarv at cesmtp.ccf.org Mon Aug 19 13:06:28 1996
From: stewarv at cesmtp.ccf.org (Valerie Stewart M.S.)

Subject: blastocyst culture -Reply
Message-ID: <s218597d.050@cesmtp.ccf.org>

Karin--

We use 5 units PMS and HCG administered at 2pm and 12pm respectively.
 Our facility is on a 12 hour cycle, 6:30am-6:30pm. I've found that
lower amounts of hormone work just as well, and sometimes better with
C57s. Also I've found that using donors selected for body weight
(13-15g at time of PMS injection) helps rather than using mice the
same age and wildly different sizes. The number of blastocysts still
varies day to day, but averages out over time. We inject 2-3 days per
clone, and get plenty of chimeras. We flush blastocysts, and I
wouldn't flush earlier than morulae--culture conditions can be
tricky.

Valerie Stewart
Transgenic/Knockout Facility
Cleveland Clinic Research Institute
"stewarv@cesmtp.ccf.org"



>>> Karin van Veen <kveen@nki.nl> - 8/15/96 6:52 AM >>>

Hi, you all!

I hope you can help me. At our institute we do zygote injections and
blastocyst injections.
For some time now we have some problems with the blastocyst yields.
It is either a superovulation problem or a flushing problem (or
both).
Can someone give me some tips concerning the time schedule on the
superovulation. In what time intervals do you inject the hormones,
and isolate the embryo's? We tried out a lot, but can not improve
our yield. We use C57/bl6 mice as donors.
We are also thinking, when it should be a flushing problem, to
isolate the zygote on day 0,5, and culture them into blastocyst to
be injected. Has anyone experience on this? How long do you have to
culture the zygotes to get blastocysts, are they good to be injected
with ES cells?
All tips are welcome!

Karin.
The Netherlands Cancer Institute
Amsterdam




From kveen at nki.nl Tue Aug 20 11:17:30 1996
From: kveen at nki.nl (Karin van Veen)

Subject: blastocyst culture THANKS
Message-ID: <Pine.SOL.3.91.960820100736.1302A-100000@Hermes.nki.nl>


Hello to everybody!

I want to thank everybody that reacted at my call for help concerning
blastocyst-yield problems.
Overall I can say that we are not doing things very wrong, most people
use more or less the same techniques and time scedules. With some minor
variations here and there. There where some tips I'm surely gonna try!
I was glad to hear that almost everybody has or had the same problems
as
we have. Blastocyst yields from B6 mice varies a lot! It goes better
and
worse, without any clear reason.
Last week I had a good week again, on three injection days, using 40
superovulated mice, I made 8 fosters, so almost 2 clones! I know, it is
not fantastic, but I was happy.
I'll let you know when I find THE answer to the problems.
Bye,
      Karin.
      The Netherlands Cancer Institute
      Amsterdam



From tjf at uci.edu Tue Aug 20 10:28:50 1996
From: tjf at uci.edu (Tom Fielder)

Subject: Pricing
Message-ID: <ae3f981e030210040088@[128.200.21.145]>

I am setting up a transgenic mouse core facility at UC-Irvine. We are
trying to decide on our recharge rates, and I was hoping to get some
feedback as to what other transgenic core facilities charge. We are
planning to offer 2 levels of service. The basic package would be a
fee
per round of injection, with no guarantees as to outcome. The deluxe
package would be a larger fee but would guarantee a certain number of
transgenic pups (maybe 2?) (with perhaps a limit on the total number of
tries). I'll try to extract as much info as I can from web pages, so
if
your prices are clearly posted (and readily accessible, e.g., through
the
Rat and Mouse Home Page, etc.), don't bother to respond. You can make
it
very brief, with just the price and what's guaranteed, and I will
summarize
the results for the list (minus names). I will post this same request
on
the Embryo Mail List and Compmed. Thanks!!
Tom




From magree00 at pop.uky.edu Wed Aug 21 10:27:02 1996
From: magree00 at pop.uky.edu (Mike Green)
Date: Wed Aug 20 11:52:52 2003
Subject: DNA quality/concentration
Message-ID: <2.2.16.19960821132702.36b743bc@pop.uky.edu>

When one of our researchers decides to produce transgenic mice, we
provide a
standard protocol for preparing the DNA for microinjection. In the past
we've had a small number of labs who have produced generally high
quality
DNA for microinjection. Now that the number of facility users is
increasing,
we're implementing a pre-injection screening of the DNA to be injected.
We
plan to run a mini-gel to verify that there is a single band of the
size and
concentration claimed by the researcher.

Do other facilities do pre-injection screening? If so, what do you look
for?

Would a spectrometer reading also be useful? Since most samples are of
low
volume and low concentration, I've been looking into the 7ul "ultra
microvolume" cells for our GeneQuant. Does anyone have experience with
these?



Mike Green                   University of Kentucky Transgenic Facility
magree00@pop.uky.edu         333 Combs
phone: 606.257.2118          800 Rose St
fax: 606.257.8940            Lexington, KY 40536
http://www.uky.edu/Transgenic/
From S.Tan at Anatomy.Unimelb.EDU.AU Thu Aug 22 14:17:47 1996
From: S.Tan at Anatomy.Unimelb.EDU.AU (Seong Seng Tan)
Date: Wed Aug 20 11:52:52 2003
Subject: DNA quality/concentration
Message-ID: <v01540b19ae41722676ae@[128.250.225.71]>

I would certainly recommend gel checking for both band size and DNA
concentration. Lately I noticed that such purified DNA when stored at
4oC
for a long time (over 12 months) are no longer able to produce
transgenic
mice by pronuclei injection. This is true even for DNA samples that
have
been proven to generated transgenic founders. Has anyone had the same
experience?


>When one of our researchers decides to produce transgenic mice, we
provide a
>standard protocol for preparing the DNA for microinjection. In the
past
>we've had a small number of labs who have produced generally high
quality
>DNA for microinjection. Now that the number of facility users is
increasing,
>we're implementing a pre-injection screening of the DNA to be
injected. We
>plan to run a mini-gel to verify that there is a single band of the
size and
>concentration claimed by the researcher.
>
>Do other facilities do pre-injection screening? If so, what do you
look for?
>
>Would a spectrometer reading also be useful? Since most samples are of
low
>volume and low concentration, I've been looking into the 7ul "ultra
>microvolume" cells for our GeneQuant. Does anyone have experience with
these?
>
>
>

Seong-Seng Tan

=======================================================================
===
Senior Lecturer, Department of Anatomy & Cell Biology
The University of Melbourne
Parkville 3052
Victoria, Australia
Fax 61 3 9 347 5219, Phone 61 3 9 344 5787
Web: http://www.anatomy.unimelb.edu.au
=======================================================================
===




From pinkert at cmed.bhs.uab.edu Thu Aug 22 07:43:11 1996
From: pinkert at cmed.bhs.uab.edu (Dr. C.A. Pinkert)
Date: Wed Aug 20 11:52:52 2003
Subject: DNA quality/concentration
Message-ID: <19960822120112826.AAA94@compaq.uab.edu>

>   Date:        Thu, 22 Aug 1996 13:17:47 +1100
>   To:          transgenic-list@ic.ac.uk
>   From:        S.Tan@Anatomy.Unimelb.EDU.AU (Seong Seng Tan)
>   Subject:     Re: DNA quality/concentration
>   Reply-to:    transgenic-list@ic.ac.uk

> I would certainly recommend gel checking for both band size and DNA
> concentration. Lately I noticed that such purified DNA when stored
at 4oC
> for a long time (over 12 months) are no longer able to produce
transgenic
> mice by pronuclei injection. This is true even for DNA samples that
have
> been proven to generated transgenic founders. Has anyone had the same
> experience?
No we haven't seen an absolute result like this. What does the gel
checking show you after a year? Is there degradation, additional
bands, or just a concentration difference?   Although we too have had
doubts about long-term storage, we routinely store aliquots at -20
and we have used an aliquot of a 'test' gene over a two year period
successfully.       C.A. Pinkert



From Allison_F._Treloar at ccmail.bms.com Thu Aug 22 09:09:07 1996
From: Allison_F._Treloar at ccmail.bms.com (Allison F. Treloar)
Date: Wed Aug 20 11:52:52 2003
Subject: DNA quality/concentration
Message-ID: <9607228407.AA840727315@ccgate0.bms.com>

>>...Lately I noticed that such purified DNA when stored at 4oC for a
long time
(over 12 months) are no longer able to produce transgenic mice by
pronuclei
injection. This is true even for DNA samples that have been proven to
generated
transgenic founders. Has anyone had the same experience?
----------------------------------------------

Absolutely! We routinely do a quick agarose check gel on stocks after
6 months
to make sure they haven't degraded (noted as multiple bands or
ladders). To
keep our stocks fresh, we also keep an aliquot frozen just in case
there is
degredation in our working solution. (Don't freeze/thaw your stocks
often,
though!) Depending on your assay, you may still get positives, even
with
degraded samples, but they won't be expressors because the whole gene
construct
probably isn't intact.

In response to S-S Tan's questions: We have the investigator send us a
gel
picture of the construct band to show a single band is indeed present.
We don't
require a spec reading, but it's a nice addition. Have the
investigator supply
it! It sure is a lot easier to have them do it than to spend money
buying
yourself new equipment.

Allison Treloar
Technical Supervisor - Transgenics
Bristol-Myers Squibb
Allison_F._Treloar@ccmail.bms.com

"I try to take one day at a time, but sometimes several days attack me
at once."




From kelly at citi2.fr Thu Aug 22 17:12:18 1996
From: kelly at citi2.fr (U344)
Date: Wed Aug 20 11:52:52 2003
Subject: 129SV Peculiarities...
Message-ID: <v02120d00ae42a5fbe787@[192.70.98.84]>

        Hello, we've got K.O. mice which are 129/bl6 hybrids, and are
trying to get our phenotypes back onto 129. Problem is, we're having a
hard time getting these animals to reproduce... very few, pregnancies,
then
very low litter yields, or many pup deaths. Does anyone have
experience
with 129SV mice who could let me know if these peculiarities are common
with this strain, or what I could do to make them happier? Any help
would
be greatly appreciated.

                                        cheers...

INSERM Unite 344 Endocrinologie Moleculaire
156 rue de Vaugirard, 75730 Paris cedex 15 France
'Man... I don't EVEN have an opinion...'




From crussell at ResGen.COM Thu Aug 22 10:23:32 1996
From: crussell at ResGen.COM (Chris Russell)
Date: Wed Aug 20 11:52:52 2003
Subject: 129SV Peculiarities...
Message-ID: <199608221416.JAA32060@twister.resgen.com>

Some info from another mouse list:


We have backcrossed a 129 ES cells derived knock-out mice into B6.
After 5
generations we ran into problems of very low fertility. We used good
B6
partners for all mating, thus the problem is not B6.

Decrease in fertility is common in intercrosses during the production
of
congenic lines but are not supposed to happen in backcrosses. During
the 5th and 6th generations of our backcross, we only obtained 1- 2
successiful litters of 4-6 animals after a year of mating each
offspring
with a healthy B6 partner.

Anyway we were able to survive the crisis, and after the 7th backcross
generation the fertility problem disappeared.

Genetic heterogeniety effects in KO mice is getting more noticed. Thus
it is
wise to carefully plan one's mating strategies or one's choice of ES
cells.



>        Hello, we've got K.O. mice which are 129/bl6 hybrids, and are
>trying to get our phenotypes back onto 129. Problem is, we're having
a
>hard time getting these animals to reproduce... very few, pregnancies,
then
>very low litter yields, or many pup deaths. Does anyone have
experience
>with 129SV mice who could let me know if these peculiarities are
common
>with this strain, or what I could do to make them happier? Any help
would
>be greatly appreciated.
>
>                                        cheers...
>
>INSERM Unite 344 Endocrinologie Moleculaire
>156 rue de Vaugirard, 75730 Paris cedex 15 France
>
>'Man... I don't EVEN have an opinion...'
>
>
>
>
>
>
>
***********************************************************************
****
                                 Christopher G. Russell, Ph.D.
                                 RESEARCH GENETICS, INC
                                     Resources for Research
-_+            _+_+           ++-_         _+-_         _+_          _++
_+-_
     -+        -+     -+       +-    +-      -+   +       -+   -+         -+    +
-+
      -+     -+       -+      +-     -+     -+    -+     -+     -+       -+
-+      -+
         -++-+         -+_-+         -+_+        -++-          -+_++         -+_+
                             2700 Memorial Parkway SW
                                    Huntsville, AL 35801
  Phone 1-800-533-4363, ext 2211 or Direct 1-800-711-2089
                                     U.K. 0-800-89-1393
                  Fax 1-205-551-1021 or 1-205-536-9016
                                    crussell@resgen.com
***********************************************************************
******




From Allison_F._Treloar at ccmail.bms.com Thu Aug 22 12:04:43 1996
From: Allison_F._Treloar at ccmail.bms.com (Allison F. Treloar)
Date: Wed Aug 20 11:52:52 2003
Subject: 129SV Peculiarities...
Message-ID: <9607228407.AA840738161@ccgate0.bms.com>

>129/bl6 hybrids....very few, pregnancies, then very low litter yields,
or many
pup deaths .

129 females are not good mothers to start off with. All the problems
you
describe above are common to this strain. I don't know how to increase
the pup
yields (superovulation?), but you can combat the pup death issue by
fostering
off the pups to better ICR moms right away. We keep a few foster moms
going for
this purpose (about 2 foster litters drop/week). Couldn't hurt ;)

Allison Treloar
Technical Supervisor-Transgenics
Bristol-Myers Squibb Co.




From tjf at uci.edu Thu Aug 22 17:33:29 1996
From: tjf at uci.edu (Tom Fielder)
Date: Wed Aug 20 11:52:52 2003
Subject: Source for M2 medium
Message-ID: <ae429eab02021004eb57@[128.200.21.145]>

Help! Does anyone know of a source for M2 medium (for manipulation of
mouse eggs) besides Sigma? They're back-ordered for 3-4 weeks. (Yes,
I
have the recipe, but not all the ingredients, nor the time to run
quality
control on home-made stuff).
Tom

Thomas J. Fielder
UC-Irvine
University Lab Animal Resources
Transgenic Mouse Facility
147 BSA
Irvine, CA 92697-1310
e-mail: tjf@uci.edu
phone: 714-824-8579
FAX: 714-824-2003




From browng at medicine.wustl.edu Fri Aug 23 07:58:44 1996
From: browng at medicine.wustl.edu (Gary Brown)
Date: Wed Aug 20 11:52:52 2003
Subject: Source for M2 medium
In-Reply-To: <ae429eab02021004eb57@[128.200.21.145]>
Message-ID: <Pine.GSO.3.94.960823065545.20926A-100000@medicine>

I think that Gibco-BRL have M2, M16, BMOC (possibly more).   Check out
this
source.

Enjoy,

Gary Brown
E-mail: 75017.1050@compuserve.com or browng@medicine.wustl.edu
Web: http://ourworld.compuserve.com/homepages/TheBroons/
              "The Microinjection Workshop"
On Thu, 22 Aug 1996, Tom Fielder wrote:

> Help! Does anyone know of a source for M2 medium (for manipulation
of
> mouse eggs) besides Sigma? They're back-ordered for 3-4 weeks.
(Yes, I
> have the recipe, but not all the ingredients, nor the time to run
quality
> control on home-made stuff).
> Tom
>
> Thomas J. Fielder
> UC-Irvine
> University Lab Animal Resources
> Transgenic Mouse Facility
> 147 BSA
> Irvine, CA 92697-1310
> e-mail: tjf@uci.edu
> phone: 714-824-8579
> FAX: 714-824-2003
>
>
>
>
>




From Allison_F._Treloar at ccmail.bms.com Fri Aug 23 09:11:00 1996
From: Allison_F._Treloar at ccmail.bms.com (Allison F. Treloar)
Date: Wed Aug 20 11:52:52 2003
Subject: Source for M2 medium
Message-ID: <9607238408.AA840814132@ccgate1.bms.com>



>Help! Does anyone know of a source for M2 medium (for manipulation of
mouse eggs) besides Sigma?

Tom,

I will be one of the hundreds of replies you'll receive who will
recommend
Specialty Media, Inc. 800.543.6029

They supply M2 (and other common media) in liquid or lyophilized powder
(my
favorite because of the longer storage time).

       liquid (50ml)   MR-015-D     $16.50
       powder (5x10ml) MR-015P-5F   $40.00

Good luck.
Allison

Technical Supervisor-Transgenics
Bristol-Myers Squibb Co.
Allison_F._Treloar@ccmail.bms.com




From mbo at avery.med.virginia.edu Fri Aug 23 09:26:07 1996
From: mbo at avery.med.virginia.edu (M. B. Ober)
Date: Wed Aug 20 11:52:52 2003
Subject: Source for M2 medium
Message-ID: <v01530500ae431b9c5927@[128.143.44.152]>

I get my M2/M16 from Specialty Media, 1-800-543-6029.   They have
ready-to-thaw and rehydratable versions.

Maggie Ober
Transgenic Mouse Core Facility
University of Virginia




From tjf at uci.edu Fri Aug 23 09:53:11 1996
From: tjf at uci.edu (Tom Fielder)
Date: Wed Aug 20 11:52:52 2003
Subject: Source for M2 medium
Message-ID: <199608231553.AA19059@e4e.oac.uci.edu>

Gary,
I had already checked the Gibco catalog and didn't see any of these
(although I didn't call). However, 4 other people have already
recommended
Specialty Media, Inc. in New Jersey (800-543-6029). Fortunately, they
have
it in stock, although at more than twice the cost of Sigma.
Tom


>I think that Gibco-BRL have M2, M16, BMOC (possibly more). Check out
this
>source.
>
>Enjoy,
>
>Gary Brown
>E-mail: 75017.1050@compuserve.com or browng@medicine.wustl.edu
>Web: http://ourworld.compuserve.com/homepages/TheBroons/
>              "The Microinjection Workshop"
>
>
>On Thu, 22 Aug 1996, Tom Fielder wrote:
>
>> Help! Does anyone know of a source for M2 medium (for manipulation
of
>> mouse eggs) besides Sigma? They're back-ordered for 3-4 weeks.
(Yes, I
>> have the recipe, but not all the ingredients, nor the time to run
quality
>> control on home-made stuff).
>> Tom
>>
>> Thomas J. Fielder
>> UC-Irvine
>> University Lab Animal Resources
>> Transgenic Mouse Facility
>> 147 BSA
>> Irvine, CA 92697-1310
>> e-mail: tjf@uci.edu
>> phone: 714-824-8579
>> FAX: 714-824-2003
>>
>>
>>
>>
>>
>
>
>
>
>
Thomas J. Fielder
UC-Irvine
University Lab Animal Resources
147 BSA
Irvine, CA 92697-1310
tjf@uci.edu




From mbo at avery.med.virginia.edu Mon Aug 26 12:45:23 1996
From: mbo at avery.med.virginia.edu (M. B. Ober)
Date: Wed Aug 20 11:52:52 2003
Subject: PMS Gonadotropin
Message-ID: <v01530501ae473aba3010@[128.143.44.152]>

Sigma has just informed me that their Pregnant Mares' Serum
Gonadotropin is
on backorder. Last time they did this it was almost 6 months before
they
had it available. What is a good alternative source?

I've been using the 50 IU/vial lyophyllized PMSG. Is this overkill?
When
Sigma had their 50IU/vial hCG on backorder I took a chance on ICN's hCG
(5000 IU for the price of one Sigma vial) and it seems to be working
fine.
Can I get away with cheaper PMSG also?

Maggie Ober
Transgenic Mouse Core Facility
University of Virginia




From sp3i at avery.med.virginia.edu Mon Aug 26 12:58:41 1996
From: sp3i at avery.med.virginia.edu (sp3i@avery.med.virginia.edu)
Date: Wed Aug 20 11:52:52 2003
Subject: PMS Gonadotropin
Message-ID: <v01540b04ae4779d0543a@[128.143.66.18]>

M
>I've been using the 50 IU/vial lyophyllized PMSG. Is this overkill?
When
>Sigma had their 50IU/vial hCG on backorder I took a chance on ICN's
hCG
>(5000 IU for the price of one Sigma vial) and it seems to be working
fine.
>Can I get away with cheaper PMSG also?


It seems there's nothing to lose by trying the cheaper PMSG if you can
get/find it.

Is it the case that 22 out of 28 hostesses were pregnant in the SM22
series?
That's my read from the database.
S

--------
Sonia Pearson-White, Ph.D.
804-982-0756 tel., 804-982-3993 fax.
University of Virginia Medical Center, MR4 Room 3002, 300 Park Place
Charlottesville, VA 22908




From PARLOW at AFP76.HUMC.EDU Mon Aug 26 10:30:50 1996
From: PARLOW at AFP76.HUMC.EDU (phone(310)222-3537 fax222-3432)
Date: Wed Aug 20 11:52:52 2003
Subject: PMS GONADOTROPIN AVAILABLE
Message-ID: <01I8Q68R7Y4I000BN6@AFP76.HUMC.EDU>

REGARDING SUPPLY OF PMSG, TWO CONCERNS ARE

1) ACCURACY OF THE STANDARDIZATION
2) COST

PMSG IS CURRENTLY AVAILABLE FROM MY LABORATORY IN A RELIABLY
STANDARDIZED FORMAT, IN ABUNDANT SUPPLY.
REQUESTS MAY BE SUBMITTED TO MY E-MAIL ADDRESS OR TO MY FAX LISTED
ABOVE OR BY MAIL TO

DR. A. F. PARLOW
HARBOR-UCLA MEDICAL CENTER
PITUITARY HORMONES/ANTISERA CENTER
1000 WEST CARSON ST.
TORRANCE, CA 90509

I WILL COME TO YOUR RESCUE.

/aFp




======= your inquiry appears below =================



From:   IN%"transgenic-list@ic.ac.uk" 26-AUG-1996 08:47:25.05
To:     IN%"transgenic-list@ic.ac.uk"
CC:
Subj:   PMS Gonadotropin

Sigma has just informed me that their Pregnant Mares' Serum
Gonadotropin is
on backorder. Last time they did this it was almost 6 months before
they
had it available. What is a good alternative source?

I've been using the 50 IU/vial lyophyllized PMSG. Is this overkill?
When
Sigma had their 50IU/vial hCG on backorder I took a chance on ICN's hCG
(5000 IU for the price of one Sigma vial) and it seems to be working
fine.
Can I get away with cheaper PMSG also?

Maggie Ober
Transgenic Mouse Core Facility
University of Virginia




From jparkert at magnus.acs.ohio-state.edu Mon Aug 26 15:55:58 1996
From: jparkert at magnus.acs.ohio-state.edu (Janice Parker-Thornburg)
Date: Wed Aug 20 11:52:52 2003
Subject: DNA quality/concentration
Message-ID: <199608261855.OAA11409@postbox.acs.ohio-state.edu>
Mike:

I guarantee a set number of transgenics with each injection, and have
found
the DNA prep to be critical. Thus, rather than trust some of the
researchers, I request a restriction digest along with a picture
proving it
is digested, and where the band of interest is circled. Then, I prep
the
DNA myself. I have had very good luck with this. No, I would not
trust
many researchers to prepare high quality DNA--especially if they have
never
done a microinjection before. As a molecular biologist myself, I know
that
one is often less than careful when prepping DNA for cloning and
digests.
However, as an injector, I also know how pure the DNA must be for this
type of work. If you can, do it yourself. I can send you an easy prep
that gives great DNA.

Jan

Jan Parker-Thornburg, Manager
Transgenic Animal Facility
The Ohio State University
jparkert@magnus.acs.ohio-state.edu




From h-marsha at nimr.mrc.ac.uk Tue Aug 27 10:42:39 1996
From: h-marsha at nimr.mrc.ac.uk (Heather)
Date: Wed Aug 20 11:52:52 2003
Subject: PMS Gonadotropin
Message-ID: <v01530500ae486c040fc6@[192.107.168.175]>

>Sigma has just informed me that their Pregnant Mares' Serum
Gonadotropin is
>on backorder. Last time they did this it was almost 6 months before
they
>had it available. What is a good alternative source?
>
>I've been using the 50 IU/vial lyophyllized PMSG. Is this overkill?
When
>Sigma had their 50IU/vial hCG on backorder I took a chance on ICN's
hCG
>(5000 IU for the price of one Sigma vial) and it seems to be working
fine.
>Can I get away with cheaper PMSG also?
>
>Maggie Ober
>Transgenic Mouse Core Facility
>University of Virginia
Hello Maggie,

This may not help you much as it involves shipping costs, but if you're
really stuck, then Intervet Labs in Cambridge, UK. may be able to help.
They provide a box of 5 vials PMS (1000IU/vial) for ?17.40. This works
out
at about ?3.50 per 1000 IU, where Sigma charge ?33.70 for the same
amount!!
Intervet's hCG is also supplied at 5 vials per box, but at 1500IU per
vial
(?38 per box). I can give you their details if you want to take this
further.

Heather Marshall
Division of Developmental Neurobiology
National Institute for Medical Research
The Ridgeway
Mill Hill
London. NW7 1AA
Tel:(0181-959-3666)

h-marsha@nimr.mrc.ac.uk




From jparkert at magnus.acs.ohio-state.edu Thu Aug 29 16:58:53 1996
From: jparkert at magnus.acs.ohio-state.edu (Janice Parker-Thornburg)
Date: Wed Aug 20 11:52:52 2003
Subject: B-cell specific expression
Message-ID: <199608291958.PAA20044@mail0.uts.ohio-state.edu>

This is just a quick query to see if I can get a response here prior to
doing some library legwork. I am working with an investigator who
wants to
have his gene expressed in B-cells in transgenic mice. I was wondering
if
anyone out there either has, or knows of someone who has a plasmid
containing the mouse IgG enhancer with an appropriate promoter and
subsequent cloning sites that has worked well in transgenics. The one
he
has now has an IgG enhancer of unknown origin followed by the promoter
for
a housekeeping gene. I'm rather skeptical that it will work to the
extent
that he needs it. I appreciate any leads.

Thanks much.

Jan Parker-Thornburg

Jan Parker-Thornburg, Manager
Transgenic Animal Facility
The Ohio State University
jparkert@magnus.acs.ohio-state.edu
From Dnxrh at aol.com Thu Aug 29 17:35:53 1996
From: Dnxrh at aol.com (Dnxrh@aol.com)
Date: Wed Aug 20 11:52:52 2003
Subject: B-cell specific expression
Message-ID: <960829163552_468124408@emout17.mail.aol.com>

You can do an on-line keyword search (without leaving your seat) of the
Oakridge National Laboratories Transgenic and Targeted Mouse Database
at:

http://www.ornl.gov/TechResources/Trans/hmepg.html

One suggestion. Once you have identified potential promotors by doing a
"B-Cell" keyword search, you might want to go back and use the promotor
itself as a keyword for a follow up search. You never know what other
tissues
a particular promotor might also express in and that can be relevant.

Rick Huntress
DNX Transgenics
dnxrh@aol.com
508-779-0189 (phone)
508-779-0190 (fax)

My opinions are my own and not necessarily that of my employer.



From Allison_F._Treloar at ccmail.bms.com Fri Aug 2 21:50:53 1996
From: Allison_F._Treloar at ccmail.bms.com (Allison F. Treloar)

Subject: Transgenic Barriers
Message-ID: <9607028390.AA839026820@ccgate1.bms.com>

Hi folks,

I am trying to get a feel for how many institutions are using barrier
conditions
for their transgenic/knockout colonies. Please reply to me off list
and I'll
sumarize for the group. Don't forget to include where you're from!

no/low barrier = conventional caging and access (+/- filter tops on
caging)

modified barrier =     access restrictions, gowning, isolators, (+/-
autoclaving)

full barrier = shower in, isolator caging, autoclaved food and bedding

Thanks in advance!
Allison Treloar
Technical Supervisor - Transgenics
Bristol-Myers Squibb

Allison_F._Treloar@ccmail.bms.com




From mgallardo at wsu.edu Fri Aug 2 21:36:49 1996
From: mgallardo at wsu.edu (Mike Gallardo)

Subject: Transgenic Barriers
Message-ID: <199608022036.NAA14361@cheetah.it.wsu.edu>

At 03:50 PM 8/2/96 -0500, you wrote:
>Hi folks,
>
>I am trying to get a feel for how many institutions are using barrier
conditions
>for their transgenic/knockout colonies. Please reply to me off list
and I'll
>sumarize for the group. Don't forget to include where you're from!
>
>no/low barrier = conventional caging and access (+/- filter tops on
caging)
>
>modified barrier = access restrictions, gowning, isolators, (+/-
autoclaving)
>
>full barrier = shower in, isolator caging, autoclaved food and bedding
>
>Thanks in advance!
>Allison Treloar
>Technical Supervisor - Transgenics
>Bristol-Myers Squibb
>
>Allison_F._Treloar@ccmail.bms.com
>
>
>Here at Wegner Hall Vivarium/WSU we use isolator caging, autoclaved
food
and bedding. We do not shower in, we use steril gowns, booties, mask,
hair
nets and gloves.
>
>

   (''`-''-/").___..--''"`-._
     `o_ o )    `-. (       ).`-.__.`)
     (_Y_.)' ._    ) `._ `. ``-..-'
   _..`--'_..- / /--' _. .'
  (il).-'' (li).'    ((!.-'     Mike Gallardo BS MS LAT
                                Administative Manager

     GO COUGS!!!!               Wegner Hall Vivarium G-44
                                Washington State University
                                Pullman, Wa 99164-6510
                                Phone (509)335-0977
                                Fax   (509)335-0162
                                e-mail mgallardo@wsu.edu
  "Brains are for thinking--pencils are for remembering"




From fmargoli at umabnet.ab.umd.edu Fri Aug 2 23:13:51 1996
From: fmargoli at umabnet.ab.umd.edu (Frank L. Margolis)

Subject: No subject
Message-ID: <v0153050fae282db061cd@[134.192.49.38]>



Anyone-

Is there an up-to-date master list of knockout and/or transgenics that
is
maintained anywhere?

Thanks

Frank L. Margolis Ph.D.
Department of Anatomy
University of Maryland at Baltimore
School of Medicine
685 West Baltimore Street
Baltimore MD 21201

Phone 410-706-8913 office
             -8914 lab
FAX          -2512 Department office




From ignelzi at umich.edu Fri Aug 2 22:15:36 1996
From: ignelzi at umich.edu (Michael A. Ignelzi, Jr.)

Subject: Transgenic Barriers
Message-ID: <v02130501ae27d9f766f8@[141.211.157.44]>

Modified barrier, access restrictions, gowning, isolators, (+/-
autoclaving), Univ of Michigan School of Dentistry (transgenic and
knockout
colonies)



>Hi folks,
>
>I am trying to get a feel for how many institutions are using barrier
>conditions
>for their transgenic/knockout colonies. Please reply to me off list
and I'll
>sumarize for the group. Don't forget to include where you're from!
>
>no/low barrier = conventional caging and access (+/- filter tops on
caging)
>
>modified barrier = access restrictions, gowning, isolators, (+/-
autoclaving)
>
>full barrier = shower in, isolator caging, autoclaved food and bedding
>
>Thanks in advance!
>Allison Treloar
>Technical Supervisor - Transgenics
>Bristol-Myers Squibb
>
>Allison_F._Treloar@ccmail.bms.com
>




From UCIVET at uci.edu Fri Aug 2 23:21:06 1996
From: UCIVET at uci.edu (Clifford R ROBERTS)

Subject: Transgenic Barriers
Message-ID: <9607028390.AA839020866@gandalf.bio.uci.edu>


low barrier: microisolator cages, changed in Class II hood, Masks, lab
coats,
gloves.

Cliff Roberts
Univ Calif, Irvine
_______________________________________________________________________
________
Subject: Transgenic Barriers
From:    transgenic-list@ic.ac.uk at biosmtp
Date:    8/2/96 1:19 PM

Hi folks,

I am trying to get a feel for how many institutions are using barrier
conditions

for their transgenic/knockout colonies. Please reply to me off list
and I'll
sumarize for the group. Don't forget to include where you're from!

no/low barrier = conventional caging and access (+/- filter tops on
caging)

modified barrier =   access restrictions, gowning, isolators, (+/-
autoclaving)
full barrier = shower in, isolator caging, autoclaved food and bedding

Thanks in advance!
Allison Treloar
Technical Supervisor - Transgenics
Bristol-Myers Squibb

Allison_F._Treloar@ccmail.bms.com




From pinkert at cmed.bhs.uab.edu Fri Aug 2 20:17:28 1996
From: pinkert at cmed.bhs.uab.edu (Dr. C.A. Pinkert)

Subject: Transgenic Barriers
Message-ID: <19960803002112601.AAD148@cap1>

> Date:           Fri, 02 Aug 96 14:21:06 PST
> To:             transgenic-list@ic.ac.uk
> Subject:        Re: Transgenic Barriers
> Reply-to:       transgenic-list@ic.ac.uk
After pulling up a number of responses to the barrier question - with
each individual describing a facility, kindly respond directly to
Allison Treloar (Allison_F._Treloar@ccmail.bms.com) as it is getting
to be a bit much.



From p.sobieszczuk at ic.ac.uk Sat Aug 3 15:53:41 1996
From: p.sobieszczuk at ic.ac.uk (Peter Sobieszczuk)

Subject: "TgList Backstage" (admin. news *1)
Message-ID: <v01510100ae28fc43f22d@[155.198.45.54]>

Dear Subscribers,

Welcome to the List, again.

The List is now about 10 days old, and quite frankly, your
administrator
(listowner) has by now much better understanding for the guys who
created
e.g. Frankenstain etc., it's easier to create the beast than to control
it.

On a positive side: The List has (as of 1st of Aug.) approximately 127
subscribers, surprisingly only, less than 40% from the old rodent-
research
list.

Congratulation to Claire and Allison for their first and second posting
(world debut on day 1 or 2 of the List existence), I will repeat their
messages below, since there were only very few people subscribed at
that
time.

On a technical note: function "reply-to" the list, was set up
deliberately
to make sending replies easier for people who are not that familiar
with
the electronics of this game, but as C.A. Pinkert noticed, this could
be
too much on occasions.

Therefore, an explanation: if you use "reply" function of your e-mail
package (e.g. "reply" from "message" menu in Eudora) remember it will
go
back to the list, not only to the original sender. To send your reply
(or
any message) just to one person use "new" message or type that address
in
the "to:" field yourself.

It would also appear to be sensible to cut out everything what is not
relevant from the copy of the original letter in your "reply to the
list"
message.

And try to remember to always fill the subject field in the new
postings as
well as signing your message (makes life easier for everyone).

Hope this will be of some help, and wishing you happy postings.


Peter
p.sobieszczuk@ic.ac.uk


P.S.

Copies of the first two posting from the last week:
____________________________________________________________________
Hello Transgenic List Subscribers:

Can anyone direct me to any studies, papers, or general information,
regarding stress indicators of mice that have been placed in the
dark or light reduced areas for 24 hours or longer ?

Thank you

Claire Smits
csmits@qlt-pdt.com
_____________________________________________________________________
I hope there are enough of you signed up on this new list to help me
out..

Aside from Genome Systems, does anyone know of other suppliers of
contract
knock-out mouse production services.   I don't know if DNX has this
service up
and running yet.

Thanks,

Allison

Technical Supervisor-Transgenics
Bristol-Myers Squibb Co.
Allison_F._Treloar@ccmail.bms.com
_____________________________________________________________________




From MEISLERM at hg-basic1mail.hg.med.umich.edu Sat Aug 3 19:04:43
1996
From: MEISLERM at hg-basic1mail.hg.med.umich.edu (MEISLERM)

Subject: brain and neuron specific promoters
Message-ID: <A145386529@HG-BASIC1MAIL.HG.MED.UMICH.EDU>

Does anyone know of a recent review of promoters with in vivo
specificity for subsets
of neurons, glia, or brain regions? We are looking for promoters
specific for brain
regions, spinal cord, motor neurons, etc.
Thanks for your suggestions.



Miriam Meisler, Ph. D.
Department of Human Genetics
University of Michigan
4708 Medical Sciences II
Ann Arbor, MI 48109-0618
tel 313 763 5546; FAX 313 763 9691.email: meislerm@umich.edu



From gstuart at UVic.CA Sun Aug 4 01:01:42 1996
From: gstuart at UVic.CA (Greg Stuart)

Subject: Rat liver anatomy question
Message-ID: <2.2.32.19960804000142.006a7bb4@pop.uvic.ca>
Hello - I'd like to draw on the collective experience of the
subscribers.
Can anybody describe where the following regions of the rat liver are
located?

- centrilobular region
- midzonal hepatocytes
- periportal regions

Also, are these regions found in each lobe of the liver, or are they
specific
to particular lobes or regions?

Please respond directly, thanks. Cheers, Greg :)

--
Greg Stuart    gstuart@uvic.ca
Centre for Environmental Health .   .*#*.            .*#*.   .*#*.
Department of Biology    * | | | * | | | *        * | | | * | | | *
University of Victoria * | | | *   * | | | *    * | | | *   * | | | *
P.O. Box 3020         * | | | *        * | | | * | | | *        * | |
|*
Victoria, B.C., CANADA `*#*'            `*#*'    `*#*'           `*#*'
V8W 3N5.
Tel (604)472-4067, Fax(604)472-4075
http://darwin.ceh.uvic.ca/students/gstuart/




From hage at med.unc.edu Sun Aug 4 03:01:14 1996
From: hage at med.unc.edu (John Hagaman)

Subject: Transgenic Barriers
Message-ID: <v01540b01ae297598a9dd@[152.2.169.3]>

>Hi folks,
>
>I am trying to get a feel for how many institutions are using barrier
>conditions
>for their transgenic/knockout colonies. Please reply to me off list
and I'll
>sumarize for the group. Don't forget to include where you're from!

we us modified barrier =   access restrictions, gowning, isolators, (+/-
autoclaving, both )

>
>Allison_F._Treloar@ccmail.bms.com
>

John R. Hagaman
Pathology Department
University of North Carolina Medical School
7525 Brinkhous-Bullitt, Room 703
Chapel Hill, N.C. 27705
966-6912 office
966-8800 Fax




From kelly at citi2.fr Mon Aug   5 23:12:58 1996
From: kelly at citi2.fr (U344)

Subject: mouse testosterone levels
Message-ID: <v02120d02ae2c09f00f80@[192.70.98.85]>

        Hello, does anyone know of a way to measure mouse testosterone
levels, or of anyone who is? I'd really like to avoid having to set up
an
ELISA for a few samples...
                                cheers...

INSERM Unite 344 Endocrinologie Moleculaire
156 rue de Vaugirard, 75730 Paris cedex 15 France

'Man... I don't EVEN have an opinion...'




From browng at medicine.wustl.edu Mon Aug 5 13:53:29 1996
From: browng at medicine.wustl.edu (Gary Brown)

Subject: WWW resource
Message-ID: <Pine.GSO.3.94.960805074227.5164A-100000@medicine>

Hi,

Just a brief wee message here from St Louis. At the risk of tooting my
own
horn I'd like to let anyone interested know about my microinjection /
transgenisis related homepage. Apologies go to those who may have seen
this on the COMPMED listings, or to those who subscribe to this list
after
my listing THIS list on the page in question! Anyways, here it is...

Page Name: The Microinjection Workshop
Page address:
http://ourworld.compuserve.com/homepages/TheBroons/homepage.htm
How best to find: Although I have registered with most of the widely
used
search engines, Yahoo is still a problem.. Best to use the Starting
Point
(www.stpt.com) and select the combination Metacrawler engine searching
on
"microinjection".
I'm trying to build up an E-mail mailing list to alert when the page is
updated (approx. 1 x per month). If interested please reply OFF-LIST
tothe
following:

75017.1050@compuserve.com

Thanx :-)

Gary




From Allison_F._Treloar at ccmail.bms.com Mon Aug 5 15:59:37 1996
From: Allison_F._Treloar at ccmail.bms.com (Allison F. Treloar)

Subject: Summary: Tg Barriers
Message-ID: <9607058392.AA839265404@ccgate1.bms.com>

First and foremost, a big thanks to everyone who responded to my
question
regarding barrier conditions for trangenic colonies. Here's the
outcome:

17 respondents
    3 facilties use low/no barrier conditions
    10 facilities use modified barrier conditions
    4 facilities use full barrier conditions


low/no barrier respondents stated that these conditions were used
primarily for
single transgenic lines or areas where investigators could generally
access
transgenic stock animals.

modified barrier users seem to rely on access restrictions, gowing,
dedicated
staff, isolator caging and autoclaved food/bedding, but don't go as far
as
shower-in or sterile clothing/scrubs. Most cage changing/animal
manipulation is
done in laminar flow hoods. This is certainly the most popular,
flexible and
cost effective option based on current opinion.

full barriers were used by a few facilities and included air showers,
sterile
clothing/scub requirements as well as all the modified barrer
conditions. No
respondents with full barriers used shower-in.

Thanks, again for all your help.

Allison
Allison_F._Treloar@ccmail.bms.com




From gina_seeburger at Merck.Com Mon Aug 5 20:07:08 1996
From: gina_seeburger at Merck.Com (Gina Seeburger)

Subject: "TgList Backstage" (admi
Message-ID: <199608052001.QAA29238@igw2>

                                                  RE>"TgList Backstage"
(admin.


--------------------------------------
Date: 8/3/96 10:56 AM
To: Gina Seeburger
From: transgenic-list@ic.ac.uk
Dear Subscribers,

Welcome to the List, again.

The List is now about 10 days old, and quite frankly, your
administrator
(listowner) has by now much better understanding for the guys who
created
e.g. Frankenstain etc., it's easier to create the beast than to control
it.

On a positive side: The List has (as of 1st of Aug.) approximately 127
subscribers, surprisingly only, less than 40% from the old rodent-
research
list.

Congratulation to Claire and Allison for their first and second posting
(world debut on day 1 or 2 of the List existence), I will repeat their
messages below, since there were only very few people subscribed at
that
time.

On a technical note: function "reply-to" the list, was set up
deliberately
to make sending replies easier for people who are not that familiar
with
the electronics of this game, but as C.A. Pinkert noticed, this could
be
too much on occasions.

Therefore, an explanation: if you use "reply" function of your e-mail
package (e.g. "reply" from "message" menu in Eudora) remember it will
go
back to the list, not only to the original sender. To send your reply
(or
any message) just to one person use "new" message or type that address
in
the "to:" field yourself.

It would also appear to be sensible to cut out everything what is not
relevant from the copy of the original letter in your "reply to the
list"
message.

And try to remember to always fill the subject field in the new
postings
as
well as signing your message (makes life easier for everyone).

Hope this will be of some help, and wishing you happy postings.


Peter
p.sobieszczuk@ic.ac.uk


P.S.

Copies of the first two posting from the last week:
____________________________________________________________________
Hello Transgenic List Subscribers:

Can anyone direct me to any studies, papers, or general information,
regarding stress indicators of mice that have been placed in the
dark or light reduced areas for 24 hours or longer ?

Thank you

Claire Smits
csmits@qlt-pdt.com
_____________________________________________________________________
I hope there are enough of you signed up on this new list to help me
out..

Aside from Genome Systems, does anyone know of other suppliers of
contract
knock-out mouse production services. I don't know if DNX has this
service
up
and running yet.

Thanks,

Allison

Technical Supervisor-Transgenics
Bristol-Myers Squibb Co.
Allison_F._Treloar@ccmail.bms.com
_____________________________________________________________________
From TSAUNDER at hg-basic1mail.hg.med.umich.edu Tue Aug 6 15:57:55
1996
From: TSAUNDER at hg-basic1mail.hg.med.umich.edu (TSAUNDER)

Subject: Meeting Announcement
Message-ID: <E62DFE6902@HG-BASIC1MAIL.HG.MED.UMICH.EDU>

THE UNIVERSITY OF MICHIGAN CENTER FOR ORGANOGENESIS
1st International Symposium

Molecular Control of Organogenesis

Saturday, October 5, 1996
Horace H. Rackham Graduate School Amphitheater
The University of Michigan
Ann Arbor, Michigan


8:45-9:00      Opening Remarks, Dr. Giles G. Bole, Dean,
   The University of Michigan Medical School Dean

9:00-9:50      Dr. Eric Olson, University of Texas, Dallas, Texas USA,
               "Genetic control of myogenesis"

9:50-10:40     Dr. Peter Gruss, Max Planck Institute of Biophysical
                            Chemistry, Germany,
  "Genes required for the development of the visual system"

10:40-10:50    Break

10:50-11:40    Dr. Cliff Tabin, Harvard Medical School, Boston,
                             Massachusetts, USA,
   "Signals patterning the vertebrate limb"

11:40-12:30    Dr. Yuh Nung Jan, University of California, San
Francisco,
                              California, USA,
    "Molecular control of neural development"

12:30-2:30     Poster Session (lunch provided)
2:30-3:20      Dr. Paul Sternberg, California Institute of Technology,
                              Pasadena, California, USA,
   "Precise formation of a hole: the C. elegans vulva"

3:20-4:10         Dr. Richard M. Harland, University of California,
Berkeley,
                                  California, USA,
   "Re-use of signaling molecules in vertebrate axis formation and
               organogenesis"

4:10-4:20         Closing Remarks, Dr. Seigo Izumo, Acting Director,
Center for
                  Organogenesis, The University of Michigan


Friday, October 4, 1996
   5:30-7:00 p.m.      Meet the Speakers at an Open Reception,
                       Museum of Art, The University of Michigan

For further information, please contact Michelle Shukait at 936-2499 or
mshukait@umich.edu




From hage at med.unc.edu Wed Aug 7 04:28:28 1996
From: hage at med.unc.edu (John Hagaman)

Subject: White noise for Mouse Embryo Yield
Message-ID: <v01540b01ae2d819d1d72@[152.2.169.1]>

Does anyone have experience re: noise in mouse rooms and how it may
affect
the yield of embryos? Do mice "get used to certain noises" Does with
drawal or introduction of noise change the yield?

Responses off list will be summarized.

Thanks

John R. Hagaman
Pathology Department
University of North Carolina Medical School
7525 Brinkhous-Bullitt, Room 703
Chapel Hill, N.C. 27705

966-6912 office
966-8800 Fax




From fmargoli at umabnet.ab.umd.edu Wed Aug 7 15:38:50 1996
From: fmargoli at umabnet.ab.umd.edu (Frank L. Margolis)

Subject: Knock out master list
Message-ID: <v01530501ae2e5aacaf10@[134.192.49.38]>

Thanks to all for your responses to my query, they were most helpful.

Frank Margolis

Frank L. Margolis Ph.D.
Department of Anatomy
University of Maryland at Baltimore
School of Medicine
685 West Baltimore Street
Baltimore MD 21201

Phone 410-706-8913 office
             -8914 lab
FAX          -2512 Department office




From r-carver at nimr.mrc.ac.uk Thu Aug 8 17:47:28 1996
From: r-carver at nimr.mrc.ac.uk (Rick Carver)

Subject: LASA Transgenics Meeting
Message-ID: <4080.9608071557@nimsn01.nimr.mrc.ac.uk>

Dear All

I am now able to give details of the speakers for the forthcoming LASA
Transgenics meeting.

Transgenesis- setting the scene               Dr   Roy Forster, Transgene
Transgenic Animals in Developmental Biology   Dr   Reter Rigby, NIMR
Transgenic Animals in Immunology              Dr   Mike Owen, ICRF
Transgenic Animals in Production Proteins     Dr   Ron James,
Pharmaceutical
Proteins Ltd
Transgenic Animals in Neuroscience            Prof Rick Lathe, Univ of
Edinburgh
Transgenic Animals in Oncology                Dr Frances Balkwell, ICRF
Transgenic Animals in Pharmaceuticals         Dr John McNeish, Pfizer
US
   Discovery and Development
Ethical and Legal Aspects of the Use of       Dr Kenneth Boyd, Univ of
Edinburgh
   Transgenic Animals


The cost will be 55 pounds stirling for LASA members and 75 pounds
stirling
for non-members. Registration forms are available from:    LASA
Speciality
Sections, PO Box 3993, Tamworth, Staffs, B78 3QU, UK. The closing date
for
registrations is 23 Aug 96.
Rick Carver
Head of Biological Services and Institute Veterinarian
National Institute for Medical Research
The Ridgeway
MILL HILL
London                     E-Mail: r-carver@nimr.mrc.ac.uk
NW7 1AA                    Tel: + 44 (0)181 959 3666 ext 2199
United Kingdom             Fax: + 44 (0)181 913 8601

"There is something fascinating about science: one gets such wholesale
returns of conjecture out of such a trifling investment of fact" - Mark
Twain




From gac at po.mri.montana.edu Thu Aug 8 00:36:49 1996
From: gac at po.mri.montana.edu (George Carlson)

Subject: discontinuation of strains
Message-ID: <v01540b06ae2ed353bf48@[204.200.84.36]>

Most of the intra-H-2 recombinant B10 congenic strains developed by the
late Dr. Jack Stimpfling of McLaughlin Research Institute will be
discontinued as live stock in the near future. Following Dr.
Stimpfling's
retirement, a grant from the American Cancer Society allowed us to
complete
isolation of his more recently identified recombinant chromosomes, to
refine localization of the crossover sites within H-2, to delimit the
congenic intervals for each strain, and to preserve the strains for
future
use. We are now in the final stage of this project and expect to have
all
strains frozen before January 1997. DNA samples will continue to be
available.

The strains are listed in:
Turner et al. Meiotic recombination within the H-2K--H-2D interval:
characterization of a panel of congenic mice, including 12 new strains,
using DNA markers. Immunogenetics 1993, 38:332-340.

Additional data on our congenic panel are being prepared for
publication.

Breeding pairs are available for any investigator who requests them. We
only ask that investigators who "adopt" a strain make it available to
others in the community and provide notification if they stop the line.

I will provide a listing of the strains that we will continue as live
stock
after we get a better idea of interest. Please let me know if you need
any
of these mice. Suggestions of other appropriate lists for posting this
notice would also be appreciated.
Thank you.

George A. Carlson (gac@po.mri.montana.edu)
McLaughlin Research Institute
1520 23rd Street South
Great Falls, Montana 59405
406 452 6208, fax: 454 6019 or 454 6004




From S.Tan at Anatomy.Unimelb.EDU.AU Thu Aug 8 09:41:53 1996
From: S.Tan at Anatomy.Unimelb.EDU.AU (Seong Seng Tan)

Subject: White noise for Mouse Embryo Yield
Message-ID: <v01540b04ae2f58725c88@[128.250.225.71]>

>Does anyone have experience re: noise in mouse rooms and how it may
affect
>the yield of embryos? Do mice "get used to certain noises" Does
with
>drawal or introduction of noise change the yield?
>

  We are currently undergoing major building renovations and this week
alone, three of our litters have been eaten by their mothers. We
suspect
loud sudden noises are responsible.

Seong-Seng Tan

=======================================================================
===
Senior Lecturer, Department of Anatomy & Cell Biology
The University of Melbourne
Parkville 3052
Victoria, Australia
Fax 61 3 9 347 5219, Phone 61 3 9 344 5787
Web: http://www.anatomy.unimelb.edu.au

=======================================================================
===




From fmargoli at umabnet.ab.umd.edu Thu Aug 8 16:11:34 1996
From: fmargoli at umabnet.ab.umd.edu (Frank L. Margolis)

Subject: Lists
Message-ID: <v01530502ae2faf1c9718@[134.192.49.38]>

To all-
In response to several requests I am posting a summary of all the info
I
have received about the availability of lists of Knockouts and
transgenics
and specific strains (without any editorial evaluation).

BioMedNet   has at least a partial list of knockouts

JAX homepage is http://www.jax.org

Current list of induced mutant strains accepted for inclusion in the
Induced Mutant Resource (IMR) at JAX is on the IMR home page-
http://lena.jax.org/resources/documents/imr/

An index of strains is available at-
http://lena.jax.org/resources/documents/imr/imr.html

and more details at-
http://lena.jax.org/resources/documents/imr/imrdata.html

for queries about any JAX mice contact-
micetech@jax.org


Also the following web sites have info that may prove to be useful-
NIH transgenic directory   Tbase
http://www.gdb.org/Dan/tbase/tbase.html

Mouse and rat research home page-
http://www.cco.caltech.edu/~mercer/htmls/rodent_page.html#homepages

and as was also pointed out to me: 1-a Medline search for info on
specific
genes or strains as well as 2- right here where I started all this.


Thanks again for all your input.

Frank Margolis

Frank L. Margolis Ph.D.
Department of Anatomy
University of Maryland at Baltimore
School of Medicine
685 West Baltimore Street
Baltimore MD 21201

Phone 410-706-8913 office
             -8914 lab
FAX          -2512 Department office




From gstuart at UVic.CA Thu Aug 8 15:37:43 1996
From: gstuart at UVic.CA (Greg Stuart)
Subject: Isolation of bone marrow & colon tissues
Message-ID: <2.2.32.19960808143743.006bd2ac@pop.uvic.ca>

Hello everyone. Does anybody have protocols that they could share for
the
isolation of rat (or rodent) bone marrow and colon tissue? Thanks, Greg
:)


--
Greg Stuart    gstuart@uvic.ca
Centre for Environmental Health .   .*#*.            .*#*.   .*#*.
Department of Biology    * | | | * | | | *        * | | | * | | | *
University of Victoria * | | | *   * | | | *    * | | | *   * | | | *
P.O. Box 3020         * | | | *        * | | | * | | | *        * | |
|*
Victoria, B.C., CANADA `*#*'            `*#*'    `*#*'           `*#*'
V8W 3N5.
Tel (604)472-4067, Fax(604)472-4075
http://darwin.ceh.uvic.ca/students/gstuart/




From CLUCK.RICHARD_S+ at LEXINGTON.VA.GOV Thu Aug 8 21:45:00 1996
From: CLUCK.RICHARD_S+ at LEXINGTON.VA.GOV (CLUCK.RICHARD_S+)

Subject: Copy of: Re: White noise for Mouse Embryo Yield
Message-ID: <4142596@LEXINGTON.VA.GOV>


You wrote:

>Does anyone have experience re: noise in mouse rooms and how it may
affect
>the yield of embryos? Do mice "get used to certain noises" Does
with
>drawal or introduction of noise change the yield?
>

       While working at the University of Kentucky, I found a number of
interuptions in breeding colonies due to the noise caused by
construction.
Liters were either canabalized by the mother or breeding stopped
completely. Hope this helps.


-----------------------------------------------------------------------
--
                                             ( )_( )
 Richard S. Cluck, B.S., LATG                    o o
 Supervisor, VMU, VA, Lexington               ==\o/==
 Research 151                                     /\    )
 Cooper Drive                                    / \ (
 Lexington, KY 40511                           /     \)
 Phone: 606-281-4927
 Fax: 606-281-4989
 E-Mail: cluck,richard_@lexington.va.gov
-----------------------------------------------------------------------
--



From r-lovell at nimr.mrc.ac.uk Fri Aug 9 17:38:45 1996
From: r-lovell at nimr.mrc.ac.uk (Robin Lovell-Badge)

Subject: Copy of: Re: White noise for Mouse Embryo Yield
Message-ID: <v01530500ae3117938cc0@[192.107.168.62]>

Dear All

There is considerable published and anecdotal stuff on noise in animal
rooms. Excessive noise, e.g. from building works, can have all sorts
of
effects which will lower reproductive efficiency. Mice seem to
particularly dislike sudden noises, so it can be beneficial to have
music
playing (classical or modern, but not too much rap !) during the day
when
cages are being changed, etc. But again, it should not be very loud,
just
enough to mask the dropping of cages, screams of students, etc.

If anyone is desperate I can try to locate published work on this - but
it
is not recent and may take a while.

Robin Lovell-badge




>You wrote:
>
>>Does anyone have experience re: noise in mouse rooms and how it may
affect
>>the yield of embryos? Do mice "get used to certain noises" Does
with
>>drawal or introduction of noise change the yield?
>>
>
>       While working at the University of Kentucky, I found a number
of
>interuptions in breeding colonies due to the noise caused by
construction.
>Liters were either canabalized by the mother or breeding stopped
>completely. Hope this helps.
>
>
>----------------------------------------------------------------------
---
>                                             ( )_( )
> Richard S. Cluck, B.S., LATG                   o o
> Supervisor, VMU, VA, Lexington               ==\o/==
> Research 151                                     /\    )
> Cooper Drive                                    / \ (
> Lexington, KY 40511                           /     \)
> Phone: 606-281-4927
> Fax: 606-281-4989
> E-Mail: cluck,richard_@lexington.va.gov
>----------------------------------------------------------------------
---




From cfoltz at MIT.EDU Mon Aug 12 11:50:40 1996
From: cfoltz at MIT.EDU (Charmaine Foltz )

Subject: control of P. pneumotropica
Message-ID: <9608121550.AA28963@MIT.MIT.EDU>

I am currently treating a small colony of Rag-2 mice to try to
eliminate
Pasteurella pneumotropica, which has been a significant pathogen for
this
particular genotype. My question to the group is how many of you have
had
experience with doing this (I am using enrofloxicin in the water).
Additionally, having read recently that humans can colonize with the
bacteria (Cudrado-Gomez LM, et al. Pasteurella pneumotropica pneumonia
in
a patient with AIDS. Clinical Inf. Dis, 1995;21:445-6), I wonder if
anyone
on the list has had experience with a barrier facility becoming
colonized
with the agent, presumably from contamination by a human source.

Thank you,
Charmaine Foltz, DVM, Dipl. ACLAM
MIT Div. Comparative Medicine
77 Massachusetts Ave. Bldg. 45
Cambridge, MA 02139
(617)252-1804
(617)258-5708 (fax)
cfoltz@mit.edu




From dknudsen at scruznet.com Tue Aug 13 04:32:45 1996
From: dknudsen at scruznet.com (Dave Knudsen)

Subject: control of P. pneumotropica
Message-ID: <v01540b01ae35a62ecf39@[165.227.113.91]>

>I am currently treating a small colony of Rag-2 mice to try to
eliminate
>Pasteurella pneumotropica, which has been a significant pathogen for
this
>particular genotype. My question to the group is how many of you have
had
>experience with doing this (I am using enrofloxicin in the water).
>Additionally, having read recently that humans can colonize with the
>bacteria (Cudrado-Gomez LM, et al. Pasteurella pneumotropica pneumonia
in
>a patient with AIDS. Clinical Inf. Dis, 1995;21:445-6), I wonder if
anyone
>on the list has had experience with a barrier facility becoming
colonized
>with the agent, presumably from contamination by a human source.

Charmaine -

I've had some experience with this agent. In a colony of over 12,000
SCID
mice used in transplantation studies (and therefore routinely
irradiated),
we were highly motivated to remove pasteurellosis from the list. Using
draconian barrier procedures and caesarian rederivation of every strain
to
be introduced into the facility, we were able to rid all of our
colonies of
P pneumo as well as everything else I know how to test for.
Individually
ventillated caging, a frequent health monitoring system, and restricted
access help kept it that way. I'll be happy to give you more details
off
line should you be interested in this approach.

Dave Knudsen DVM, MS, DACLAM
Scotts Valley, California, USA
<dknudsen@scruznet.com>

***
"In my judgement it is either an enigma or some kind of bug. If it
dies, I
will take it apart and see what its arrangements are. I never had a
thing
perplex me so." Mark Twain, 1893 (from "Extracts from Adam's Diary")
***




From dbowtell at petermac.unimelb.edu.au Tue Aug 13 11:49:47 1996
From: dbowtell at petermac.unimelb.edu.au (David Bowtell)

Subject: Double KO with increased G418
Message-ID: <v01530501ae3699bc24e4@[203.4.164.101]>

We have used increased G418 concentration as a means of selecting
homozgous
cells on two occassions. One construct had a pGKNeo casssette and the
cells
were still laughing in 4mg/ml (we blinked first). The second construct
was
promoterless, the cells were marginally G418 resistant and it worked
like a
dream. The paper by Mortensen (MCB 12, 2391) used a pGKNeo cassette
but I
am unclear whether it was the point mutant (low activity) neo cassette.
So
we would be interested in other people's experience with this approach
to
double KO in ES cells (and any tidbits on experience targeting the
second
allele indepndantly).

Many thanks.

David Bowtell.

******* Please note new email address **********
David Bowtell
Principal Research Fellow
Research Division
Peter MacCallum Cancer Institute
Locked Bag 1 A'Beckett St,
Melbourne 3000

Lab 61-3-96561287
Office 61-3-96561296
Fax 61-3-96561411
email: dbowtell@petermac.unimelb.edu.au




From wixson at biovax.dnet.basf-ag.de Tue Aug 13 18:16:55 1996
From: wixson at biovax.dnet.basf-ag.de (Sally Wixson, VMD, MS)

Subject: Knockout advice
Message-ID: <9608131716.AA23717@gwr.zxd.basf-ag.de>

    I direct the Lab Animal department of a pharmaceutical research
company
that has an active, but disorganized, knock out team. I have been here
eight
months and trying to get their animal work back on track. I would be
grateful
for your advice on several topics:
.....what strain of vactomized males do you use and why? How do you
assess
their sterility (other than ordering then vasec. by the vendor)? How
often do
you use vasec. males? (my invesitagors use each male once/week or
less!)
.....what strain of embryo recipients do you use for ES cell work? Our
donors
are C57BL/6, recipients are B6D2F1's. My team wants a "big" target so
they keep
the females here three months or so on a high fat diet so they are big
enough
to inject. Obviously, this leads to a waste of cage space. Many people
use
ICR's or CD1's as recipients because they are a big target and good
moms!

.....in breeding chimeras, our PI insists that each chimera must
produce100
pups (black) before being considered a failure at germ line
transmission. Many
of the chimeras are lowly sterile or sterile, so this format entails
breeding
formany months to more than a year. Several other labs told me their
chimeras
get three matings and if no agouti pups are produced, they are ENA'd.

.....do you produce your own ES cells or get them from
donation/purchase from
other labs? We are currently buying the RW4 ES cell line from Genome
Systems.
Do you require MAP testing of the new ES cell lines?

.....do you superovulate your females for embryo donation? My PI says
this doe
not yield viable embryos for ES cell work, so they breed 4-5X the
number of
females they rpedict will be needed for each injectionso that they have
enough
embryos.

Many thanks for the advice,
Sally K, Wixson, VMD, MS



From wixson at biovax.dnet.basf-ag.de Tue Aug 13 18:16:55 1996
From: wixson at biovax.dnet.basf-ag.de (Sally Wixson, VMD, MS)

Subject: Knockout advice
Message-ID: <9608131716.AA23717@gwr.zxd.basf-ag.de>

    I direct the Lab Animal department of a pharmaceutical research
company
that has an active, but disorganized, knock out team. I have been here
eight
months and trying to get their animal work back on track. I would be
grateful
for your advice on several topics:
.....what strain of vactomized males do you use and why? How do you
assess
their sterility (other than ordering then vasec. by the vendor)? How
often do
you use vasec. males? (my invesitagors use each male once/week or
less!)

.....what strain of embryo recipients do you use for ES cell work? Our
donors
are C57BL/6, recipients are B6D2F1's. My team wants a "big" target so
they keep
the females here three months or so on a high fat diet so they are big
enough
to inject. Obviously, this leads to a waste of cage space. Many people
use
ICR's or CD1's as recipients because they are a big target and good
moms!

.....in breeding chimeras, our PI insists that each chimera must
produce100
pups (black) before being considered a failure at germ line
transmission. Many
of the chimeras are lowly sterile or sterile, so this format entails
breeding
formany months to more than a year. Several other labs told me their
chimeras
get three matings and if no agouti pups are produced, they are ENA'd.

.....do you produce your own ES cells or get them from
donation/purchase from
other labs? We are currently buying the RW4 ES cell line from Genome
Systems.
Do you require MAP testing of the new ES cell lines?

.....do you superovulate your females for embryo donation? My PI says
this doe
not yield viable embryos for ES cell work, so they breed 4-5X the
number of
females they rpedict will be needed for each injectionso that they have
enough
embryos.

Many thanks for the advice,
Sally K, Wixson, VMD, MS



From dbowtell at petermac.unimelb.edu.au Tue Aug 13 11:49:47 1996
From: dbowtell at petermac.unimelb.edu.au (David Bowtell)

Subject: Double KO with increased G418
Message-ID: <v01530501ae3699bc24e4@[203.4.164.101]>

We have used increased G418 concentration as a means of selecting
homozgous
cells on two occassions. One construct had a pGKNeo casssette and the
cells
were still laughing in 4mg/ml (we blinked first). The second construct
was
promoterless, the cells were marginally G418 resistant and it worked
like a
dream. The paper by Mortensen (MCB 12, 2391) used a pGKNeo cassette
but I
am unclear whether it was the point mutant (low activity) neo cassette.
So
we would be interested in other people's experience with this approach
to
double KO in ES cells (and any tidbits on experience targeting the
second
allele indepndantly).

Many thanks.

David Bowtell.

******* Please note new email address **********
David Bowtell
Principal Research Fellow
Research Division
Peter MacCallum Cancer Institute
Locked Bag 1 A'Beckett St,
Melbourne 3000

Lab 61-3-96561287
Office 61-3-96561296
Fax 61-3-96561411
email: dbowtell@petermac.unimelb.edu.au




From dknudsen at scruznet.com Tue Aug 13 04:32:45 1996
From: dknudsen at scruznet.com (Dave Knudsen)

Subject: control of P. pneumotropica
Message-ID: <v01540b01ae35a62ecf39@[165.227.113.91]>

>I am currently treating a small colony of Rag-2 mice to try to
eliminate
>Pasteurella pneumotropica, which has been a significant pathogen for
this
>particular genotype. My question to the group is how many of you have
had
>experience with doing this (I am using enrofloxicin in the water).
>Additionally, having read recently that humans can colonize with the
>bacteria (Cudrado-Gomez LM, et al. Pasteurella pneumotropica pneumonia
in
>a patient with AIDS. Clinical Inf. Dis, 1995;21:445-6), I wonder if
anyone
>on the list has had experience with a barrier facility becoming
colonized
>with the agent, presumably from contamination by a human source.

Charmaine -

I've had some experience with this agent. In a colony of over 12,000
SCID
mice used in transplantation studies (and therefore routinely
irradiated),
we were highly motivated to remove pasteurellosis from the list. Using
draconian barrier procedures and caesarian rederivation of every strain
to
be introduced into the facility, we were able to rid all of our
colonies of
P pneumo as well as everything else I know how to test for.
Individually
ventillated caging, a frequent health monitoring system, and restricted
access help kept it that way. I'll be happy to give you more details
off
line should you be interested in this approach.

Dave Knudsen DVM, MS, DACLAM
Scotts Valley, California, USA
<dknudsen@scruznet.com>

***
"In my judgement it is either an enigma or some kind of bug. If it
dies, I
will take it apart and see what its arrangements are. I never had a
thing
perplex me so." Mark Twain, 1893 (from "Extracts from Adam's Diary")
***




From cfoltz at MIT.EDU Mon Aug 12 11:50:40 1996
From: cfoltz at MIT.EDU (Charmaine Foltz )

Subject: control of P. pneumotropica
Message-ID: <9608121550.AA28963@MIT.MIT.EDU>

I am currently treating a small colony of Rag-2 mice to try to
eliminate
Pasteurella pneumotropica, which has been a significant pathogen for
this
particular genotype. My question to the group is how many of you have
had
experience with doing this (I am using enrofloxicin in the water).
Additionally, having read recently that humans can colonize with the
bacteria (Cudrado-Gomez LM, et al. Pasteurella pneumotropica pneumonia
in
a patient with AIDS. Clinical Inf. Dis, 1995;21:445-6), I wonder if
anyone
on the list has had experience with a barrier facility becoming
colonized
with the agent, presumably from contamination by a human source.

Thank you,
Charmaine Foltz, DVM, Dipl. ACLAM
MIT Div. Comparative Medicine
77 Massachusetts Ave. Bldg. 45
Cambridge, MA 02139
(617)252-1804
(617)258-5708 (fax)
cfoltz@mit.edu




From browng at medicine.wustl.edu Wed Aug 14 13:52:59 1996
From: browng at medicine.wustl.edu (Gary Brown)

Subject: Knockout advice
In-Reply-To: <9608131716.AA23717@gwr.zxd.basf-ag.de>
Message-ID: <Pine.GSO.3.94.960814071607.1135A-100000@medicine>

Hi,

Just throwing in my 2 cents worth...

On Tue, 13 Aug 1996, Sally Wixson, VMD, MS wrote:

> .....what strain of vactomized males do you use and why? How do you
assess
> their sterility (other than ordering then vasec. by the vendor)? How
often do
> you use vasec. males? (my invesitagors use each male once/week or
less!)

I use Swiss Webster VAS-X males, ordered in from Taconic Farms. In my
previous position we did our own vasectomies on C57Bl/6J males
(pronuclear
microinjection only) and assessed their sterility by co-housing with
Swiss
Webster (or other inexpensive white mouse strain) females (2) for
approx.
6 weeks prior to use. Males were vasectomized at 4-5 weeks of age,
excising a minimum of 1cm of vas deferens per side.
 >
> .....what strain of embryo recipients do you use for ES cell work?
Our donors
> are C57BL/6, recipients are B6D2F1's. My team wants a "big" target so
they keep
> the females here three months or so on a high fat diet so they are
big enough
> to inject. Obviously, this leads to a waste of cage space. Many
people use
> ICR's or CD1's as recipients because they are a big target and good
moms!

I have used B6D2F1s for recipients although now use Swiss Webster
(occasionally ICR) females as they are a relatively large mouse strain.
As for feeding them on a high fat diet I personally do not subscribe to
the 'big target' philosophy - at least no bigger than using regular
mouse
chow for my chosen strain. Since we have a pool of SW females from
which
to pick those in estrus and individuals may be passed over several
times,
high fat diet pushes some of them to a size where surgery becomes more
difficult due to the increased fat deposits around the oviduct/ovary/UT
junction, and I believe that the increased incision size in the body
wall
that this necessitates increases the risk to the animal.
 >
> .....in breeding chimeras, our PI insists that each chimera must
produce100
> pups (black) before being considered a failure at germ line
transmission. Many
> of the chimeras are lowly sterile or sterile, so this format entails
breeding
> formany months to more than a year. Several other labs told me their
chimeras
> get three matings and if no agouti pups are produced, they are ENA'd.

This is insane! My initial reaction would be to tell your PI to get a
life! Our policy is to discard all female chimeras (low germline
transmission, increased sterility in the F1 generation when successful)
and to discard all >50% (by color coat) male chimeras. 3 matings
(strikes) and you're out sounds appropriate to me.
 >
> .....do you produce your own ES cells or get them from
donation/purchase from
> other labs? We are currently buying the RW4 ES cell line from Genome
Systems.
> Do you require MAP testing of the new ES cell lines?

Our lab has approached other labs for ES cells, although we are trying
to
establish our own cell line currently. With reference to the RW4 cell
line, I have used this effectively on several occasions and note that
it
has a high color coat conversion rate. I also know the individual who
created this cell line, and can forward any questions you may have to
her.
Our institution requires MAP testing of all ES cell lines for use in
mice.
>
> .....do you superovulate your females for embryo donation? My PI says
this doe
> not yield viable embryos for ES cell work, so they breed 4-5X the
number of
> females they rpedict will be needed for each injectionso that they
have enough
> embryos.
>
Yes, I superovulate females for embryo donation. I typically
superovulate
7 C57Bl/6J females for one day's effort, with a variation between 20-25
morula per plugged female. I culture these in the incubator overnight
to
blastocysts with an 85-95% conversion rate. The females used are
between
3.5-4.5 weeks old. I use Sigma Chemical Co. new embryo tested PMSG and
HCG. My stud males are mated no more than once weekly. There is a
reference for superovulating females for blastocyst production, but I
would have to look it up on Medline.   Sounds to me like your PI needs
to
do some homework!

Hope this helps!


Gary Brown
E-mail: 75017.1050@compuserve.com or browng@medicine.wustl.edu
Web: http://ourworld.compuserve.com/homepages/TheBroons/
                "The Microinjection Workshop"




From kveen at nki.nl Thu Aug 15 11:52:09 1996
From: kveen at nki.nl (Karin van Veen)

Subject: blastocyst culture
Message-ID: <Pine.SOL.3.91.960815113939.13618B-100000@Hermes.nki.nl>


Hi, you all!

I hope you can help me. At our institute we do zygote injections and
blastocyst injections.
For some time now we have some problems with the blastocyst yields. It
is
either a superovulation problem or a flushing problem (or both).
Can someone give me some tips concerning the time schedule on the
superovulation. In what time intervals do you inject the hormones, and
isolate the embryo's? We tried out a lot, but can not improve our
yield.
We use C57/bl6 mice as donors.
We are also thinking, when it should be a flushing problem, to isolate
the
zygote on day 0,5, and culture them into blastocyst to be injected. Has
anyone experience on this? How long do you have to culture the zygotes
to get
blastocysts, are they good to be injected with ES cells?
All tips are welcome!

Karin.
The Netherlands Cancer Institute
Amsterdam




From browng at medicine.wustl.edu Thu Aug 15 13:23:30 1996
From: browng at medicine.wustl.edu (Gary Brown)

Subject: blastocyst culture
In-Reply-To: <Pine.SOL.3.91.960815113939.13618B-100000@Hermes.nki.nl>
Message-ID: <Pine.GSO.3.94.960815070704.14243A-100000@medicine>
On Thu, 15 Aug 1996, Karin van Veen wrote:
> Can someone give me some tips concerning the time schedule on the
> superovulation. In what time intervals do you inject the hormones,
and
> isolate the embryo's?
> We use C57/bl6 mice as donors.

We use a 12hr/12hr light-dark cycle from 4am to 4pm in our facility,
coupled with 47hrs between PMSG and HCG. These hormones are typically
given at 10.30am and (47hrs later) 9.30am repectively for this light
cycle. We use hormones obtained through Sigma Chemical Co. (Embryo
culture tested - new product) and have been obtaining yields of MORULAE
at
a rate of 20-25 per plugged C57Bl/6J female. These morulae are
cultured
overnight to blastocysts for ES cell injection in Gibco's modified
BMOC-3
culture media with pen/strep.added in a 37deg C 5% CO2 incubator.

> We are also thinking, when it should be a flushing problem, to
isolate the
> zygote on day 0,5, and culture them into blastocyst to be injected.
Has
> anyone experience on this? How long do you have to culture the
zygotes to get
> blastocysts, are they good to be injected with ES cells?
> All tips are welcome!
>
For culture past day 0.5, you would need 3 more days and some
consistently
good media. I suggest you flush morulae! Although I doubt that
flushing
would present too much of a problem, this is how I do it. Using a
short
bevel stainless 30g hypo., introduce the needle at the last oviduct
loop
before the UT junction and flush media through this area expelling the
embryos out of the uterus (cut 1cm down from the oviduct). If this is
problematical, flush through the infundibulum (requires a little more
finesse).

Hope this helps out!

Gary Brown
Email: browng@medicine.wustl.edu or 75017.1050@compuserve.com
Web: http://ourworld.compuserve.com/homepages/TheBroons/
              "The Microinjection Workshop"




From m.hampson at ic.ac.uk Thu Aug 15 17:18:40 1996
From: m.hampson at ic.ac.uk (m.hampson@ic.ac.uk)

Subject: Test - please ignore
Message-ID: <E0ur584-0001l4-00@sphinx>
Test - please ignore

--
     +-------------------------------------------------------------------
-+
     | Martyn Hampson          |    Tel:    0171 594 6973
|
     | Imperial College        |    Fax:    0171 594 6958
|
     | Computer Centre         |    E-Mail: M.Hampson@ic.ac.uk
|
     | London SW7 2BP, ENGLAND |    "Don't just do something, sit there!"
|
     +-------------------------------------------------------------------
-+




From m.hampson at ic.ac.uk Thu Aug 15 17:26:16 1996
From: m.hampson at ic.ac.uk (m.hampson@ic.ac.uk)

Subject: Test please ignore
Message-ID: <E0ur5FQ-0001lc-00@sphinx>

Test please ignore
--
   +-------------------------------------------------------------------
-+
   | Martyn Hampson          |    Tel:    0171 594 6973
|
   | Imperial College        |    Fax:    0171 594 6958
|
   | Computer Centre         |    E-Mail: M.Hampson@ic.ac.uk
|
   | London SW7 2BP, ENGLAND |    "Don't just do something, sit there!"
|
   +-------------------------------------------------------------------
-+




From m.hampson at ic.ac.uk Thu Aug 15 17:29:17 1996
From: m.hampson at ic.ac.uk (m.hampson@ic.ac.uk)

Subject: Test - please ignore
Message-ID: <E0ur5IL-0001lj-00@sphinx>



 Test - please ignore
--
   +-------------------------------------------------------------------
-+
     | Martyn Hampson          |    Tel:    0171 594 6973
|
     | Imperial College        |    Fax:    0171 594 6958
|
     | Computer Centre         |    E-Mail: M.Hampson@ic.ac.uk
|
     | London SW7 2BP, ENGLAND |    "Don't just do something, sit there!"
|
     +-------------------------------------------------------------------
-+




From m.hampson at ic.ac.uk Thu Aug 15 17:31:25 1996
From: m.hampson at ic.ac.uk (m.hampson@ic.ac.uk)

Subject: Test - please ignore
Message-ID: <E0ur5KP-0001lr-00@sphinx>

Test - please ignore
--
   +-------------------------------------------------------------------
-+
   | Martyn Hampson          |    Tel:    0171 594 6973
|
   | Imperial College        |    Fax:    0171 594 6958
|
   | Computer Centre         |    E-Mail: M.Hampson@ic.ac.uk
|
   | London SW7 2BP, ENGLAND |    "Don't just do something, sit there!"
|
   +-------------------------------------------------------------------
-+




From gina_seeburger at Merck.Com Thu Aug 15 21:15:46 1996
From: gina_seeburger at Merck.Com (Gina Seeburger)

Subject: blastocyst culture
Message-ID: <199608160406.AAA22680@igw2.merck.com>

                                      REPLY o REPLY o REPLY o REPLY
                                      RE>blastocyst culture
Karin,
We were having problems too here in the US, but giving the first
injection(PMSG, Sigma brand) between 3:00pm and 4 (.3cc, .7.5IU) and
the
second (hCG) around 1:00 seems to help. We are also using C57BL/6
donors
for blast injections.   Collected around 7:30am. Also summer seems to
create a low.
Good luck
Gina Seeburger
Merck & Co,Inc.
gina_seeburger@merck.com

--------------------------------------
Date: 8/15/96 5:57 AM
To: Gina Seeburger
From: transgenic-list@ic.ac.uk

Hi, you all!

I hope you can help me. At our institute we do zygote injections and
blastocyst injections.
For some time now we have some problems with the blastocyst yields. It
is
either a superovulation problem or a flushing problem (or both).
Can someone give me some tips concerning the time schedule on the
superovulation. In what time intervals do you inject the hormones, and
isolate the embryo's? We tried out a lot, but can not improve our
yield.
We use C57/bl6 mice as donors.
We are also thinking, when it should be a flushing problem, to isolate
the

zygote on day 0,5, and culture them into blastocyst to be injected. Has
anyone experience on this? How long do you have to culture the zygotes
to
get
blastocysts, are they good to be injected with ES cells?
All tips are welcome!

Karin.
The Netherlands Cancer Institute
Amsterdam




From tjf at uci.edu Fri Aug 16 20:11:55 1996
From: tjf at uci.edu (Tom Fielder)

Subject: oocyte injection
Message-ID: <ae3a74350a021004d502@[128.200.21.145]>

        I am brand new to the field of transgenics, and I am setting up
a
transgenic core facility at UC-Irvine. I am still trying to optimize
my
pipets for injection of DNA into oocytes. I'm using a Sutter P-97
puller,
and 1mm OD x 0.78mm ID capillaries with internal filament. I got some
pipets that seemed to work very well yesterday, judging by the swelling
of
the pronucleus and no visible cell contents leaking out after
withdrawing
the pipet, but this morning 65% of the injected cells had died
(cytoplasm
was condensed and dark brown), while none of the uninjected controls
looked
like this. I'm afraid that the taper of the needle is too abrupt near
the
tip (i.e., it gets too big too fast). A very patient person at Sutter
is
helping me with the pipets, but my question for the list is this: Is
the
condensed, dark brown cytoplasm indicative of mechanical trauma, or is
it a
result of bad DNA/buffer, or something else?
        A second question: The books recommend using females that are
24-25 days old for superovulation. What happens if you use older ones?
Will the yield of oocytes be significantly less? I've been using 6-8
week
old FVB mice and getting about 30 eggs/mouse, but I've noticed that a
large
number of them aren't fertilized, even from the females that were
obviously
plugged. Any thoughts?
Tom

Thomas J. Fielder
UC-Irvine
University Lab Animal Resources
Transgenic Mouse Facility
147 BSA
Irvine, CA 92697-1310
e-mail: tjf@uci.edu
phone: 714-824-8579
FAX: 714-824-2003




From MKENNEDY at chmeds.ac.nz Sat Aug 17 11:12:42 1996
From: MKENNEDY at chmeds.ac.nz (MKENNEDY@chmeds.ac.nz)

Subject: Double KO with increased G418
Message-ID: <960817221242.c621@chmeds.ac.nz>

Date sent:   17-AUG-1996 22:06:32

Hi Dave,

Just got back from this year's Queenstown Meeting, which (for my
sins) I organised.

>We have used increased G418 concentration as a means of selecting
homozgous
>cells on two occassions. One construct had a pGKNeo casssette and the
cells
>were still laughing in 4mg/ml (we blinked first). The second
construct was
>promoterless, the cells were marginally G418 resistant and it worked
like a
>dream. The paper by Mortensen (MCB 12, 2391) used a pGKNeo cassette
but I
>am unclear whether it was the point mutant (low activity) neo
cassette. So
>we would be interested in other people's experience with this approach
to
>double KO in ES cells (and any tidbits on experience targeting the
second
>allele indepndantly).

For what it is worth, I sequenced Mortensen's pNTK construct, which
I am pretty sure was the one he used in the MCB paper, and it was
of the mutant neo variety. I am at home at present, but if you
haven't seen it, then John Sedivy had a very interesting MCB a
year or so back on titrating G418 to obtain high efficiency KO's,
with some interesting observations that related to the homozygous
KO method. Let me know if you need more info, and I'll dig it
out when I am at work. I have an oligo from close to the
polymorphic site in the neo gene, if you want to check other neo
constructs by sequencing - you're welcome to some of it.

Cheers,

Martin

NNNN   NN    Martin A Kennedy (E-mail = mkennedy@chmeds.ac.nz)   ZZZZZZZ
NN NN NN          Cytogenetic and Molecular Oncology Unit           ZZZ
NN NN NN              Christchurch School of Medicine             ZZZ
NN   NNNN                Christchurch, New Zealand               ZZZZZZZ
                 Phone (64-3)364-0880   Fax (64-3)364-0750



From stewarv at cesmtp.ccf.org Mon Aug 19 16:40:44 1996
From: stewarv at cesmtp.ccf.org (Valerie Stewart M.S.)

Subject: Knockout advice -Reply
Message-ID: <s2185375.030@cesmtp.ccf.org>

Hi Sally--

We use Swiss Webster vasectomized males, bought from Taconic. We
don't routinely test them, and have so far (knock on wood!) never had
a leak. When I used to do the vasectomies myself, we tested them with
two females each, let them plug both, then waited 3 weeks for
hopefully no resultant pregnancies. We use vasectomized males twice
a week to every other day. They can be used every night for a couple
of weeks before needing some down time. Once a week is ridiculous,
they don't need to keep their sperm count up after all! We use them
for a year as well, or until they stop performing. We use Swiss
Webster or CD-1 females for recipients--they are easy to estrus
select, good mothers, and cheap. I wouldn't keep them on a high fat
diet--in fact, we routinely cull out recipients that are too fat and
discard them.
As for testing germline transmission, 100 pups to "prove" lack of
transmission is the standard, but practicality usually dicatates
compromise. We only breed males over 50% chimeric, females only if
there are no male chimeras available. Three litters is a good
number, if they're good-sized litters, I usually try for 25-30 pups.

We are using E14.1 cells from Cincinnati, but I've used CCE, D3, J1,
R1, etc. Cell line is crucial; I've heard Genome Systems' line works
fine.

We superovulate for blastocysts routinely, using C57Bl6 females. We
generally do 10 at a time, with the standard amounts/timing of
hormones, and a 6:30am/6:30pm light cycle. The one thing we find is
crucial is to match body weight of the donors, 13-15g at time of PMS
administration. We do three injection days per clone, averaging 30
blastocysts per 10 donors (with the occasional null day). I have
gotten good results with number and % chimerism, as well as germline
transmission with these blastocysts. We only use the males once per
week, and usually only for 6-8 months, though a colleague says she
uses them twice a week for a year with no trouble.

Good Luck--
Valerie Stewart
Transgenic/Knockout Facility
Cleveland Clinic Research Institute
"stewarv@cesmtp.ccf.org"

>>> Sally Wixson, VMD, MS <wixson@biovax.dnet.basf-ag.de> - 8/13/96
1:16 PM >>>
    I direct the Lab Animal department of a pharmaceutical research
company that has an active, but disorganized, knock out team. I have
been here eight months and trying to get their animal work back on
track. I would be grateful for your advice on several topics:
.....what strain of vactomized males do you use and why? How do you
assess their sterility (other than ordering then vasec. by the
vendor)? How often do you use vasec. males? (my invesitagors use
each male once/week or less!)

.....what strain of embryo recipients do you use for ES cell work?
Our donors are C57BL/6, recipients are B6D2F1's. My team wants a
"big" target so they keep the females here three months or so on a
high fat diet so they are big enough to inject. Obviously, this
leads to a waste of cage space. Many people use ICR's or CD1's as
recipients because they are a big target and good moms!

.....in breeding chimeras, our PI insists that each chimera must
produce100 pups (black) before being considered a failure at germ
line transmission. Many of the chimeras are lowly sterile or
sterile, so this format entails breeding formany months to more than
a year. Several other labs told me their chimeras get three matings
and if no agouti pups are produced, they are ENA'd.

.....do you produce your own ES cells or get them from
donation/purchase from other labs? We are currently buying the RW4
ES cell line from Genome Systems. Do you require MAP testing of the
new ES cell lines?
.....do you superovulate your females for embryo donation? My PI says
this doe not yield viable embryos for ES cell work, so they breed
4-5X the number of females they rpedict will be needed for each
injectionso that they have enough embryos.

Many thanks for the advice,
Sally K, Wixson, VMD, MS




From crussell at ResGen.COM Mon Aug 19 17:03:24 1996
From: crussell at ResGen.COM (Chris Russell)

Subject: employment opportunity
Message-ID: <199608191556.KAA21398@twister.resgen.com>

Position Available Immediately!
Lab Animal Technologist/Research Assistant

Skills Required:
--Knowledgeable in theoretical embryology
--Skilled in applied embryology and microsurgery techniques
--Primarily laboratory work using the mouse as a model system
--Molecular biology experience preferred

--Salary: $28-30K plus great      benefits

--Excellent work environment in a growing biotech firm in Huntsville,
AL

Please send resume and references to:
Dr. Chris Russell
Research Genetics, Inc.
2700 Memorial Parkway, SW
Huntsville, AL 35801
1-800-711-2089
e-mail: crussell@resgen.com
***********************************************************************
****
                                 Christopher G. Russell, Ph.D.
                                 RESEARCH GENETICS, INC
                                     Resources for Research
-_+            _+_+           ++-_         _+-_         _+_          _++
_+-_
     -+        -+     -+       +-    +-      -+   +       -+   -+         -+    +
-+
      -+     -+       -+      +-     -+     -+    -+     -+     -+       -+
-+      -+
         -++-+         -+_-+         -+_+        -++-          -+_++         -+_+
                             2700 Memorial Parkway SW
                                    Huntsville, AL 35801
 Phone 1-800-533-4363, ext 2211 or Direct 1-800-711-2089
                                     U.K. 0-800-89-1393
                  Fax 1-205-551-1021 or 1-205-536-9016
                              crussell@resgen.com
***********************************************************************
******




From stewarv at cesmtp.ccf.org Mon Aug 19 17:06:28 1996
From: stewarv at cesmtp.ccf.org (Valerie Stewart M.S.)

Subject: blastocyst culture -Reply
Message-ID: <s218597d.050@cesmtp.ccf.org>

Karin--

We use 5 units PMS and HCG administered at 2pm and 12pm respectively.
 Our facility is on a 12 hour cycle, 6:30am-6:30pm. I've found that
lower amounts of hormone work just as well, and sometimes better with
C57s. Also I've found that using donors selected for body weight
(13-15g at time of PMS injection) helps rather than using mice the
same age and wildly different sizes. The number of blastocysts still
varies day to day, but averages out over time. We inject 2-3 days per
clone, and get plenty of chimeras. We flush blastocysts, and I
wouldn't flush earlier than morulae--culture conditions can be
tricky.

Valerie Stewart
Transgenic/Knockout Facility
Cleveland Clinic Research Institute
"stewarv@cesmtp.ccf.org"



>>> Karin van Veen <kveen@nki.nl> - 8/15/96 6:52 AM >>>

Hi, you all!

I hope you can help me. At our institute we do zygote injections and
blastocyst injections.
For some time now we have some problems with the blastocyst yields.
It is either a superovulation problem or a flushing problem (or
both).
Can someone give me some tips concerning the time schedule on the
superovulation. In what time intervals do you inject the hormones,
and isolate the embryo's? We tried out a lot, but can not improve
our yield. We use C57/bl6 mice as donors.
We are also thinking, when it should be a flushing problem, to
isolate the zygote on day 0,5, and culture them into blastocyst to
be injected. Has anyone experience on this? How long do you have to
culture the zygotes to get blastocysts, are they good to be injected
with ES cells?
All tips are welcome!

Karin.
The Netherlands Cancer Institute
Amsterdam
From kveen at nki.nl Tue Aug 20 10:17:30 1996
From: kveen at nki.nl (Karin van Veen)

Subject: blastocyst culture THANKS
Message-ID: <Pine.SOL.3.91.960820100736.1302A-100000@Hermes.nki.nl>


Hello to everybody!

I want to thank everybody that reacted at my call for help concerning
blastocyst-yield problems.
Overall I can say that we are not doing things very wrong, most people
use more or less the same techniques and time scedules. With some minor
variations here and there. There where some tips I'm surely gonna try!
I was glad to hear that almost everybody has or had the same problems
as
we have. Blastocyst yields from B6 mice varies a lot! It goes better
and
worse, without any clear reason.
Last week I had a good week again, on three injection days, using 40
superovulated mice, I made 8 fosters, so almost 2 clones! I know, it is
not fantastic, but I was happy.
I'll let you know when I find THE answer to the problems.
Bye,
      Karin.
      The Netherlands Cancer Institute
      Amsterdam



From tjf at uci.edu Tue Aug 20 17:28:50 1996
From: tjf at uci.edu (Tom Fielder)

Subject: Pricing
Message-ID: <ae3f981e030210040088@[128.200.21.145]>

I am setting up a transgenic mouse core facility at UC-Irvine. We are
trying to decide on our recharge rates, and I was hoping to get some
feedback as to what other transgenic core facilities charge. We are
planning to offer 2 levels of service. The basic package would be a
fee
per round of injection, with no guarantees as to outcome. The deluxe
package would be a larger fee but would guarantee a certain number of
transgenic pups (maybe 2?) (with perhaps a limit on the total number of
tries). I'll try to extract as much info as I can from web pages, so
if
your prices are clearly posted (and readily accessible, e.g., through
the
Rat and Mouse Home Page, etc.), don't bother to respond. You can make
it
very brief, with just the price and what's guaranteed, and I will
summarize
the results for the list (minus names). I will post this same request
on
the Embryo Mail List and Compmed. Thanks!!
Tom




From magree00 at pop.uky.edu Wed Aug 21 14:27:02 1996
From: magree00 at pop.uky.edu (Mike Green)

Subject: DNA quality/concentration
Message-ID: <2.2.16.19960821132702.36b743bc@pop.uky.edu>

When one of our researchers decides to produce transgenic mice, we
provide a
standard protocol for preparing the DNA for microinjection. In the past
we've had a small number of labs who have produced generally high
quality
DNA for microinjection. Now that the number of facility users is
increasing,
we're implementing a pre-injection screening of the DNA to be injected.
We
plan to run a mini-gel to verify that there is a single band of the
size and
concentration claimed by the researcher.

Do other facilities do pre-injection screening? If so, what do you look
for?

Would a spectrometer reading also be useful? Since most samples are of
low
volume and low concentration, I've been looking into the 7ul "ultra
microvolume" cells for our GeneQuant. Does anyone have experience with
these?



Mike Green                   University of Kentucky Transgenic Facility
magree00@pop.uky.edu         333 Combs
phone: 606.257.2118          800 Rose St
fax: 606.257.8940            Lexington, KY 40536
http://www.uky.edu/Transgenic/




From S.Tan at Anatomy.Unimelb.EDU.AU Thu Aug 22 03:17:47 1996
From: S.Tan at Anatomy.Unimelb.EDU.AU (Seong Seng Tan)

Subject: DNA quality/concentration
Message-ID: <v01540b19ae41722676ae@[128.250.225.71]>

I would certainly recommend gel checking for both band size and DNA
concentration. Lately I noticed that such purified DNA when stored at
4oC
for a long time (over 12 months) are no longer able to produce
transgenic
mice by pronuclei injection. This is true even for DNA samples that
have
been proven to generated transgenic founders. Has anyone had the same
experience?


>When one of our researchers decides to produce transgenic mice, we
provide a
>standard protocol for preparing the DNA for microinjection. In the
past
>we've had a small number of labs who have produced generally high
quality
>DNA for microinjection. Now that the number of facility users is
increasing,
>we're implementing a pre-injection screening of the DNA to be
injected. We
>plan to run a mini-gel to verify that there is a single band of the
size and
>concentration claimed by the researcher.
>
>Do other facilities do pre-injection screening? If so, what do you
look for?
>
>Would a spectrometer reading also be useful? Since most samples are of
low
>volume and low concentration, I've been looking into the 7ul "ultra
>microvolume" cells for our GeneQuant. Does anyone have experience with
these?
>
>
>

Seong-Seng Tan

=======================================================================
===
Senior Lecturer, Department of Anatomy & Cell Biology
The University of Melbourne
Parkville 3052
Victoria, Australia
Fax 61 3 9 347 5219, Phone 61 3 9 344 5787
Web: http://www.anatomy.unimelb.edu.au

=======================================================================
===




From pinkert at cmed.bhs.uab.edu Thu Aug 22 07:43:11 1996
From: pinkert at cmed.bhs.uab.edu (Dr. C.A. Pinkert)
Subject: DNA quality/concentration
Message-ID: <19960822120112826.AAA94@compaq.uab.edu>

>   Date:        Thu, 22 Aug 1996 13:17:47 +1100
>   To:          transgenic-list@ic.ac.uk
>   From:        S.Tan@Anatomy.Unimelb.EDU.AU (Seong Seng Tan)
>   Subject:     Re: DNA quality/concentration
>   Reply-to:    transgenic-list@ic.ac.uk

> I would certainly recommend gel checking for both band size and DNA
> concentration. Lately I noticed that such purified DNA when stored
at 4oC
> for a long time (over 12 months) are no longer able to produce
transgenic
> mice by pronuclei injection. This is true even for DNA samples that
have
> been proven to generated transgenic founders. Has anyone had the same
> experience?
No we haven't seen an absolute result like this. What does the gel
checking show you after a year? Is there degradation, additional
bands, or just a concentration difference?   Although we too have had
doubts about long-term storage, we routinely store aliquots at -20
and we have used an aliquot of a 'test' gene over a two year period
successfully.       C.A. Pinkert



From Allison_F._Treloar at ccmail.bms.com Thu Aug 22 14:09:07 1996
From: Allison_F._Treloar at ccmail.bms.com (Allison F. Treloar)

Subject: DNA quality/concentration
Message-ID: <9607228407.AA840727315@ccgate0.bms.com>

>>...Lately I noticed that such purified DNA when stored at 4oC for a
long time
(over 12 months) are no longer able to produce transgenic mice by
pronuclei
injection. This is true even for DNA samples that have been proven to
generated
transgenic founders. Has anyone had the same experience?
----------------------------------------------

Absolutely! We routinely do a quick agarose check gel on stocks after
6 months
to make sure they haven't degraded (noted as multiple bands or
ladders). To
keep our stocks fresh, we also keep an aliquot frozen just in case
there is
degredation in our working solution. (Don't freeze/thaw your stocks
often,
though!) Depending on your assay, you may still get positives, even
with
degraded samples, but they won't be expressors because the whole gene
construct
probably isn't intact.
In response to S-S Tan's questions: We have the investigator send us a
gel
picture of the construct band to show a single band is indeed present.
We don't
require a spec reading, but it's a nice addition. Have the
investigator supply
it! It sure is a lot easier to have them do it than to spend money
buying
yourself new equipment.

Allison Treloar
Technical Supervisor - Transgenics
Bristol-Myers Squibb
Allison_F._Treloar@ccmail.bms.com

"I try to take one day at a time, but sometimes several days attack me
at once."




From kelly at citi2.fr Fri Aug 23 01:12:18 1996
From: kelly at citi2.fr (U344)

Subject: 129SV Peculiarities...
Message-ID: <v02120d00ae42a5fbe787@[192.70.98.84]>

        Hello, we've got K.O. mice which are 129/bl6 hybrids, and are
trying to get our phenotypes back onto 129. Problem is, we're having a
hard time getting these animals to reproduce... very few, pregnancies,
then
very low litter yields, or many pup deaths. Does anyone have
experience
with 129SV mice who could let me know if these peculiarities are common
with this strain, or what I could do to make them happier? Any help
would
be greatly appreciated.

                                           cheers...

INSERM Unite 344 Endocrinologie Moleculaire
156 rue de Vaugirard, 75730 Paris cedex 15 France

'Man... I don't EVEN have an opinion...'




From crussell at ResGen.COM   Thu Aug 22 15:23:32 1996
From: crussell at ResGen.COM (Chris Russell)

Subject: 129SV Peculiarities...
Message-ID: <199608221416.JAA32060@twister.resgen.com>

Some info from another mouse list:


We have backcrossed a 129 ES cells derived knock-out mice into B6.
After 5
generations we ran into problems of very low fertility. We used good
B6
partners for all mating, thus the problem is not B6.

Decrease in fertility is common in intercrosses during the production
of
congenic lines but are not supposed to happen in backcrosses. During
the 5th and 6th generations of our backcross, we only obtained 1- 2
successiful litters of 4-6 animals after a year of mating each
offspring
with a healthy B6 partner.

Anyway we were able to survive the crisis, and after the 7th backcross
generation the fertility problem disappeared.

Genetic heterogeniety effects in KO mice is getting more noticed. Thus
it is
wise to carefully plan one's mating strategies or one's choice of ES
cells.



>        Hello, we've got K.O. mice which are 129/bl6 hybrids, and are
>trying to get our phenotypes back onto 129. Problem is, we're having
a
>hard time getting these animals to reproduce... very few, pregnancies,
then
>very low litter yields, or many pup deaths. Does anyone have
experience
>with 129SV mice who could let me know if these peculiarities are
common
>with this strain, or what I could do to make them happier? Any help
would
>be greatly appreciated.
>
>                                         cheers...
>
>INSERM Unite 344 Endocrinologie Moleculaire
>156 rue de Vaugirard, 75730 Paris cedex 15 France
>
>'Man... I don't EVEN have an opinion...'
>
>
>
>
>
>
>
***********************************************************************
****
                                 Christopher G. Russell, Ph.D.
                                 RESEARCH GENETICS, INC
                                     Resources for Research
-_+            _+_+           ++-_         _+-_         _+_          _++
_+-_
     -+        -+     -+       +-    +-      -+   +       -+   -+         -+    +
-+
      -+     -+       -+      +-     -+     -+    -+     -+     -+       -+
-+      -+
         -++-+         -+_-+         -+_+        -++-          -+_++         -+_+
                             2700 Memorial Parkway SW
                                    Huntsville, AL 35801
  Phone 1-800-533-4363, ext 2211 or Direct 1-800-711-2089
                                     U.K. 0-800-89-1393
                  Fax 1-205-551-1021 or 1-205-536-9016
                                    crussell@resgen.com
***********************************************************************
******




From Allison_F._Treloar at ccmail.bms.com Thu Aug 22 17:04:43 1996
From: Allison_F._Treloar at ccmail.bms.com (Allison F. Treloar)

Subject: 129SV Peculiarities...
Message-ID: <9607228407.AA840738161@ccgate0.bms.com>

>129/bl6 hybrids....very few, pregnancies, then very low litter yields,
or many
pup deaths .

129 females are not good mothers to start off with. All the problems
you
describe above are common to this strain. I don't know how to increase
the pup
yields (superovulation?), but you can combat the pup death issue by
fostering
off the pups to better ICR moms right away. We keep a few foster moms
going for
this purpose (about 2 foster litters drop/week). Couldn't hurt ;)

Allison Treloar
Technical Supervisor-Transgenics
Bristol-Myers Squibb Co.
From tjf at uci.edu Fri Aug 23 00:33:29 1996
From: tjf at uci.edu (Tom Fielder)

Subject: Source for M2 medium
Message-ID: <ae429eab02021004eb57@[128.200.21.145]>

Help! Does anyone know of a source for M2 medium (for manipulation of
mouse eggs) besides Sigma? They're back-ordered for 3-4 weeks. (Yes,
I
have the recipe, but not all the ingredients, nor the time to run
quality
control on home-made stuff).
Tom

Thomas J. Fielder
UC-Irvine
University Lab Animal Resources
Transgenic Mouse Facility
147 BSA
Irvine, CA 92697-1310
e-mail: tjf@uci.edu
phone: 714-824-8579
FAX: 714-824-2003




From browng at medicine.wustl.edu Fri Aug 23 12:58:44 1996
From: browng at medicine.wustl.edu (Gary Brown)

Subject: Source for M2 medium
In-Reply-To: <ae429eab02021004eb57@[128.200.21.145]>
Message-ID: <Pine.GSO.3.94.960823065545.20926A-100000@medicine>

I think that Gibco-BRL have M2, M16, BMOC (possibly more).   Check out
this
source.

Enjoy,

Gary Brown
E-mail: 75017.1050@compuserve.com or browng@medicine.wustl.edu
Web: http://ourworld.compuserve.com/homepages/TheBroons/
              "The Microinjection Workshop"


On Thu, 22 Aug 1996, Tom Fielder wrote:

> Help! Does anyone know of a source for M2 medium (for manipulation
of
> mouse eggs) besides Sigma? They're back-ordered for 3-4 weeks.
(Yes, I
> have the recipe, but not all the ingredients, nor the time to run
quality
>   control on home-made stuff).
>   Tom
>
>   Thomas J. Fielder
>   UC-Irvine
>   University Lab Animal Resources
>   Transgenic Mouse Facility
>   147 BSA
>   Irvine, CA 92697-1310
>   e-mail: tjf@uci.edu
>   phone: 714-824-8579
>   FAX: 714-824-2003
>
>
>
>
>




From Allison_F._Treloar at ccmail.bms.com Fri Aug 23 14:11:00 1996
From: Allison_F._Treloar at ccmail.bms.com (Allison F. Treloar)

Subject: Source for M2 medium
Message-ID: <9607238408.AA840814132@ccgate1.bms.com>



>Help! Does anyone know of a source for M2 medium (for manipulation of
mouse eggs) besides Sigma?

Tom,

I will be one of the hundreds of replies you'll receive who will
recommend
Specialty Media, Inc. 800.543.6029

They supply M2 (and other common media) in liquid or lyophilized powder
(my
favorite because of the longer storage time).

       liquid (50ml)   MR-015-D     $16.50
       powder (5x10ml) MR-015P-5F   $40.00

Good luck.

Allison

Technical Supervisor-Transgenics
Bristol-Myers Squibb Co.
Allison_F._Treloar@ccmail.bms.com
From mbo at avery.med.virginia.edu Fri Aug 23 13:26:07 1996
From: mbo at avery.med.virginia.edu (M. B. Ober)

Subject: Source for M2 medium
Message-ID: <v01530500ae431b9c5927@[128.143.44.152]>

I get my M2/M16 from Specialty Media, 1-800-543-6029.   They have
ready-to-thaw and rehydratable versions.

Maggie Ober
Transgenic Mouse Core Facility
University of Virginia




From tjf at uci.edu Fri Aug 23 16:53:11 1996
From: tjf at uci.edu (Tom Fielder)

Subject: Source for M2 medium
Message-ID: <199608231553.AA19059@e4e.oac.uci.edu>

Gary,
I had already checked the Gibco catalog and didn't see any of these
(although I didn't call). However, 4 other people have already
recommended
Specialty Media, Inc. in New Jersey (800-543-6029). Fortunately, they
have
it in stock, although at more than twice the cost of Sigma.
Tom


>I think that Gibco-BRL have M2, M16, BMOC (possibly more). Check out
this
>source.
>
>Enjoy,
>
>Gary Brown
>E-mail: 75017.1050@compuserve.com or browng@medicine.wustl.edu
>Web: http://ourworld.compuserve.com/homepages/TheBroons/
>              "The Microinjection Workshop"
>
>
>On Thu, 22 Aug 1996, Tom Fielder wrote:
>
>> Help! Does anyone know of a source for M2 medium (for manipulation
of
>> mouse eggs) besides Sigma? They're back-ordered for 3-4 weeks.
(Yes, I
>> have the recipe, but not all the ingredients, nor the time to run
quality
>> control on home-made stuff).
>> Tom
>>
>> Thomas J. Fielder
>> UC-Irvine
>> University Lab Animal Resources
>> Transgenic Mouse Facility
>> 147 BSA
>> Irvine, CA 92697-1310
>> e-mail: tjf@uci.edu
>> phone: 714-824-8579
>> FAX: 714-824-2003
>>
>>
>>
>>
>>
>
>
>
>
>
Thomas J. Fielder
UC-Irvine
University Lab Animal Resources
147 BSA
Irvine, CA 92697-1310
tjf@uci.edu




From mbo at avery.med.virginia.edu Mon Aug 26 16:45:23 1996
From: mbo at avery.med.virginia.edu (M. B. Ober)

Subject: PMS Gonadotropin
Message-ID: <v01530501ae473aba3010@[128.143.44.152]>

Sigma has just informed me that their Pregnant Mares' Serum
Gonadotropin is
on backorder. Last time they did this it was almost 6 months before
they
had it available. What is a good alternative source?

I've been using the 50 IU/vial lyophyllized PMSG. Is this overkill?
When
Sigma had their 50IU/vial hCG on backorder I took a chance on ICN's hCG
(5000 IU for the price of one Sigma vial) and it seems to be working
fine.
Can I get away with cheaper PMSG also?

Maggie Ober
Transgenic Mouse Core Facility
University of Virginia
From sp3i at avery.med.virginia.edu Mon Aug 26 16:58:41 1996
From: sp3i at avery.med.virginia.edu (sp3i@avery.med.virginia.edu)

Subject: PMS Gonadotropin
Message-ID: <v01540b04ae4779d0543a@[128.143.66.18]>

M
>I've been using the 50 IU/vial lyophyllized PMSG. Is this overkill?
When
>Sigma had their 50IU/vial hCG on backorder I took a chance on ICN's
hCG
>(5000 IU for the price of one Sigma vial) and it seems to be working
fine.
>Can I get away with cheaper PMSG also?


It seems there's nothing to lose by trying the cheaper PMSG if you can
get/find it.

Is it the case that 22 out of 28 hostesses were pregnant in the SM22
series?
That's my read from the database.
S

--------
Sonia Pearson-White, Ph.D.
804-982-0756 tel., 804-982-3993 fax.
University of Virginia Medical Center, MR4 Room 3002, 300 Park Place
Charlottesville, VA 22908




From PARLOW at AFP76.HUMC.EDU Mon Aug 26 17:30:50 1996
From: PARLOW at AFP76.HUMC.EDU (phone(310)222-3537 fax222-3432)

Subject: PMS GONADOTROPIN AVAILABLE
Message-ID: <01I8Q68R7Y4I000BN6@AFP76.HUMC.EDU>

REGARDING SUPPLY OF PMSG, TWO CONCERNS ARE

1) ACCURACY OF THE STANDARDIZATION
2) COST

PMSG IS CURRENTLY AVAILABLE FROM MY LABORATORY IN A RELIABLY
STANDARDIZED FORMAT, IN ABUNDANT SUPPLY.

REQUESTS MAY BE SUBMITTED TO MY E-MAIL ADDRESS OR TO MY FAX LISTED
ABOVE OR BY MAIL TO

DR. A. F. PARLOW
HARBOR-UCLA MEDICAL CENTER
PITUITARY HORMONES/ANTISERA CENTER
1000 WEST CARSON ST.
TORRANCE, CA 90509
I WILL COME TO YOUR RESCUE.

/aFp




======= your inquiry appears below =================



From:   IN%"transgenic-list@ic.ac.uk" 26-AUG-1996 08:47:25.05
To:     IN%"transgenic-list@ic.ac.uk"
CC:
Subj:   PMS Gonadotropin

Sigma has just informed me that their Pregnant Mares' Serum
Gonadotropin is
on backorder. Last time they did this it was almost 6 months before
they
had it available. What is a good alternative source?

I've been using the 50 IU/vial lyophyllized PMSG. Is this overkill?
When
Sigma had their 50IU/vial hCG on backorder I took a chance on ICN's hCG
(5000 IU for the price of one Sigma vial) and it seems to be working
fine.
Can I get away with cheaper PMSG also?

Maggie Ober
Transgenic Mouse Core Facility
University of Virginia




From jparkert at magnus.acs.ohio-state.edu Mon Aug 26 19:55:58 1996
From: jparkert at magnus.acs.ohio-state.edu (Janice Parker-Thornburg)

Subject: DNA quality/concentration
Message-ID: <199608261855.OAA11409@postbox.acs.ohio-state.edu>

Mike:

I guarantee a set number of transgenics with each injection, and have
found
the DNA prep to be critical. Thus, rather than trust some of the
researchers, I request a restriction digest along with a picture
proving it
is digested, and where the band of interest is circled. Then, I prep
the
DNA myself. I have had very good luck with this. No, I would not
trust
many researchers to prepare high quality DNA--especially if they have
never
done a microinjection before. As a molecular biologist myself, I know
that
one is often less than careful when prepping DNA for cloning and
digests.
However, as an injector, I also know how pure the DNA must be for this
type of work. If you can, do it yourself. I can send you an easy prep
that gives great DNA.

Jan

Jan Parker-Thornburg, Manager
Transgenic Animal Facility
The Ohio State University
jparkert@magnus.acs.ohio-state.edu




From h-marsha at nimr.mrc.ac.uk Tue Aug 27 09:42:39 1996
From: h-marsha at nimr.mrc.ac.uk (Heather)

Subject: PMS Gonadotropin
Message-ID: <v01530500ae486c040fc6@[192.107.168.175]>

>Sigma has just informed me that their Pregnant Mares' Serum
Gonadotropin is
>on backorder. Last time they did this it was almost 6 months before
they
>had it available. What is a good alternative source?
>
>I've been using the 50 IU/vial lyophyllized PMSG. Is this overkill?
When
>Sigma had their 50IU/vial hCG on backorder I took a chance on ICN's
hCG
>(5000 IU for the price of one Sigma vial) and it seems to be working
fine.
>Can I get away with cheaper PMSG also?
>
>Maggie Ober
>Transgenic Mouse Core Facility
>University of Virginia


Hello Maggie,

This may not help you much as it involves shipping costs, but if you're
really stuck, then Intervet Labs in Cambridge, UK. may be able to help.
They provide a box of 5 vials PMS (1000IU/vial) for ?17.40. This works
out
at about ?3.50 per 1000 IU, where Sigma charge ?33.70 for the same
amount!!
Intervet's hCG is also supplied at 5 vials per box, but at 1500IU per
vial
(?38 per box). I can give you their details if you want to take this
further.

Heather Marshall
Division of Developmental Neurobiology
National Institute for Medical Research
The Ridgeway
Mill Hill
London. NW7 1AA
Tel:(0181-959-3666)

h-marsha@nimr.mrc.ac.uk




From jparkert at magnus.acs.ohio-state.edu Thu Aug 29 20:58:53 1996
From: jparkert at magnus.acs.ohio-state.edu (Janice Parker-Thornburg)

Subject: B-cell specific expression
Message-ID: <199608291958.PAA20044@mail0.uts.ohio-state.edu>

This is just a quick query to see if I can get a response here prior to
doing some library legwork. I am working with an investigator who
wants to
have his gene expressed in B-cells in transgenic mice. I was wondering
if
anyone out there either has, or knows of someone who has a plasmid
containing the mouse IgG enhancer with an appropriate promoter and
subsequent cloning sites that has worked well in transgenics. The one
he
has now has an IgG enhancer of unknown origin followed by the promoter
for
a housekeeping gene. I'm rather skeptical that it will work to the
extent
that he needs it. I appreciate any leads.

Thanks much.

Jan Parker-Thornburg

Jan Parker-Thornburg, Manager
Transgenic Animal Facility
The Ohio State University
jparkert@magnus.acs.ohio-state.edu




From Dnxrh at aol.com Thu Aug 29 21:35:53 1996
From: Dnxrh at aol.com (Dnxrh@aol.com)

Subject: B-cell specific expression
Message-ID: <960829163552_468124408@emout17.mail.aol.com>

You can do an on-line keyword search (without leaving your seat) of the
Oakridge National Laboratories Transgenic and Targeted Mouse Database
at:

http://www.ornl.gov/TechResources/Trans/hmepg.html

One suggestion. Once you have identified potential promotors by doing a
"B-Cell" keyword search, you might want to go back and use the promotor
itself as a keyword for a follow up search. You never know what other
tissues
a particular promotor might also express in and that can be relevant.

Rick Huntress
DNX Transgenics
dnxrh@aol.com
508-779-0189 (phone)
508-779-0190 (fax)

My opinions are my own and not necessarily that of my employer.




1996 September

From DAMAK at whio.lincoln.ac.nz Mon Sep 2 14:32:54 1996
From: DAMAK at whio.lincoln.ac.nz (Damak, Sami)
Date: Wed Aug 20 11:52:52 2003
Subject: Import of transgenic mice into the US
Message-ID: <17CA2DC2C3E@whio.lincoln.ac.nz>

I am looking at sending transgenic mice from New Zealand to the US.
Does anyone know who I should approach to get customs and quarantine
clearance?
Thanks
Sami Damak



From Dnxrh at aol.com Tue Sep 3 00:07:29 1996
From: Dnxrh at aol.com (Dnxrh@aol.com)
Date: Wed Aug 20 11:52:52 2003
Subject: Import of transgenic mice into the US (DNA Too!)
Message-ID: <960902230729_275590968@emout12.mail.aol.com>

If permits are required (they may not be) it is the responsibility of
the
importer (your US recipient) to get the permits. Have them contact the
USDA -
APHIS to determine if permits are needed.
USDA - APHIS can be found at:
http://www.aphis.usda.gov
301-734-7885 (phone)
301-734-8226

The guidelines seem to change but at the present time you should know
the
following:

Do the mice contain any livestock or avian genes ?

Do the mice contain and genes from any livestock or avian pathogens?

Are the mice infected with any known diseases which are transferable to
humans, birds or livestock?

Do the mice contain any genes from human pathogens?

Have the mice been treated with any human blood products or any
livestock
derived products (ie. albumin).

If you can answer "no" to these questions then your recipient in the US
will
probably will not need a permit. Even if a permit is not required, the
animals will still need to have paperwork documenting where they come
from,
what genes they contain and what their current health status is.

If you do need a permit they can take 6 weeks to get.

I have had more experience shipping mice out of the US than in shipping
them
into the US. I suggest very strongly that you or your recipient in the
US
contact USDA-APHIS directly to confirm this information.

PS. to all you gene jockeys out there. These same rules apply to all
recombinant DNA as well. We have had plasmids and DNA fragments held up
by
customs because the invoices did not state clearly all of the above.

Good Luck,
Rick Huntress
DNX Transgenics
dnxrh@aol.com
+508-779-0189 (phone)
+508-779-0190 (fax)

My opinions are my own and not necessarily that of my employer.




From WRIGHTA at gunet.georgetown.edu Tue Sep 3 12:07:11 1996
From: WRIGHTA at gunet.georgetown.edu (Ann Wright)
Date: Wed Aug 20 11:52:52 2003
Subject: Import of transgenic mice into the US -Reply
Message-ID: <s22c13c8.006@gunet.georgetown.edu>

Information regarding APHIS programs and import-export permits can be
obtained on the home page http://www.aphis.usda.gov. The fax number
is 301-734-8226, email ncie@aphis.usda.gov or voice mail
301-734-4412. The permit to import animals is needed to clear customs.




From carton at murray.fordham.edu Tue Sep 3 20:02:12 1996
From: carton at murray.fordham.edu (Jill Carton)
Date: Wed Aug 20 11:52:52 2003
Subject: electroporators for ES cells
Message-ID: <v02130500ae5263ad7834@[169.132.99.88]>

I am a graduate student at Fordham University working on generating a
knockout mouse. The electroporation protocols I have found for the ES
cells are all using the Biorad Gene Pulser. We have in our department
a
Baekon 2000 gene transfer system. I was wondering if anyone has ever
used
this apparatus for electroporation of ES cells and if you would share
your
protocol with me. This would save me a lot of time trying to work out
the
conditions.

Thanks in advance for the information,
              Jill Carton

Jill Carton
Dept. Biological Sciences
Fordham University
Larkin Hall, rm 160
Bronx, NY 10458
718-817-3652
718-817-3645 (fax)
carton@murray.fordham.edu




From jklohse at facstaff.wisc.edu Fri Sep 6 18:57:44 1996
From: jklohse at facstaff.wisc.edu (Jan K. Lohse)
Date: Wed Aug 20 11:52:52 2003
Subject: transgene control systems
Message-ID: <v02140b01ae565caab1f7@[144.92.48.83]>


I'm interested in finding out what sorts of transgenic mice people are
currently creating with non-leaky, induceable promoter systems such as
the
ones that use the tet operon. Are these being used with good success?
Satisfactory expression, turned on or off by a drug as designed? Is
the
lac operon being used for this sort of thing as well?   Any
descriptions of
constructs, suggestions, and info about potential problems you're
willing
to share would be appreciated.

Also, how about similar info about recombination modification systems
such
as CRE-LOX and FLP?

Any other cool control systems out there?

Thanks very much.   I think many of us will find the answers
interesting.

Jan




From fmargoli at umabnet.ab.umd.edu Mon Sep 9 10:36:33 1996
From: fmargoli at umabnet.ab.umd.edu (Frank L. Margolis)
Date: Wed Aug 20 11:52:52 2003
Subject: transgene control systems
Message-ID: <v01530500ae59db81c04d@[134.192.49.38]>

>I'm interested in finding out what sorts of transgenic mice people are
>currently creating with non-leaky, induceable promoter systems such as
the
>ones that use the tet operon. Are these being used with good success?
>Satisfactory expression, turned on or off by a drug as designed? Is
the
>lac operon being used for this sort of thing as well?   Any
descriptions of
>constructs, suggestions, and info about potential problems you're
willing
>to share would be appreciated.
>
>Also, how about similar info about recombination modification systems
such
>as CRE-LOX and FLP?
>
>Any other cool control systems out there?
>
>Thanks very much. I think many of us will find the answers
interesting.
>
>Jan
Similar questions arise with regard to the ecdysone regulated promoter
recently reported in PNAS. Any info would be useful to all.

Frank Margolis

Frank L. Margolis Ph.D.
Department of Anatomy
University of Maryland at Baltimore
School of Medicine
685 West Baltimore Street
Baltimore MD 21201

Phone 410-706-8913 office
             -8914 lab
FAX          -2512 Department office

"There is something fascinating about science: one gets such wholesale
returns of conjecture out of such a trifling investment of fact."
Mark
Twain




From Roger.Leemann at imr.psi.ch Mon Sep 9 20:37:54 1996
From: Roger.Leemann at imr.psi.ch (Roger C. Leemann)
Date: Wed Aug 20 11:52:52 2003
Subject: visibility of cell nuclei in tw-cell mouse embryos
Message-ID: <323453844d7e002@pss200.psi.ch>

Hi

Gary Brown came across a few questions I had sent to the rodent-
research
list and because he had to admit that his answers to most of them would
be
"I don't know" suggested I might give it a try on this list.

Sure, I'd like to, although it's only indirectly related to transgenic
mice. Thanks Gary.

I use 2-cell embryos of B6C3F1xB6C3F1 (Charles River, Germany) in a
project
where we want to study the effects of irradiation (e.g. alpha, protons)
on
the level of individual cells. We are able to detect single particle
tracks
with a track detector (basically a photographic emulsion) which is
irradiated together with the cells or embryos with a vertical beam
(normal
to the plane of the detector).
With a microscope we take pictures of both the cells or embryos and the
developed detector and superimpose the two images. To tell which cell
nuclei have been hit by a particle we must of course be able to see the
boundary of the nucleus (we would like to get away without fluorescent
staining).
I have always had difficulties observing the nucleus in brightfield or
phase contrast (we don't have Nomarski and Hoffman is no solution) in
my
B6C3F1 embryos. By chance I found out that in an other mouse strain
(NMRI)
the nucleus is clearly visible even with bright field illumination.
Unfortunatly the NMRI embryos didn't develop beyond the 4-cell stage
under
my plain vanilla culture conditions which are OK for the B6C3F1.

My question therefore: Is anybody aware of a mouse strain, where the
nuclei
of life two-cell embryos are easily visible (unstained, no Nomarksi)
and
which is healthy enough, so that the embryos will develop in vitro to
the
blastocyst stage and hatch under standard culture conditions?

Any help is highly appreciated.

Roger

:::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::
:::::
Work: Institute for Medical Radiobiology        ++41 (56) 310-3792
[desk]
       CH-5232 Villigen PSI                     ++41 (1) 3856-561
[desk ZH]
       Switzerland                              ++41 (56) 310 3294
[fax]
Home: Nordstrasse 26, CH-8006 Zuerich
       Switzerland                              ++41 (1) 361 0349
[home]




From pinkert at cmed.bhs.uab.edu Mon Sep 9 13:56:56 1996
From: pinkert at cmed.bhs.uab.edu (Dr. C.A. Pinkert)
Date: Wed Aug 20 11:52:52 2003
Subject: visibility of cell nuclei in tw-cell mouse embryos
Message-ID: <19960909180142886.AAA77@cap1>

>   Date:        Mon, 9 Sep 1996 19:37:54 +0200
>   To:          transgenic-list@ic.ac.uk
>   From:        Roger.Leemann@imr.psi.ch (Roger C. Leemann)
>   Subject:     visibility of cell nuclei in tw-cell mouse embryos
>   Reply-to:    transgenic-list@ic.ac.uk

> Hi
>
> Gary Brown came across a few questions I had sent to the rodent-
research
> list and because he had to admit that his answers to most of them
would be
> "I don't know" suggested I might give it a try on this list.
>
> Sure, I'd like to, although it's only indirectly related to
transgenic
> mice. Thanks Gary.
>
> I use 2-cell embryos of B6C3F1xB6C3F1 (Charles River, Germany) in a
project
> where we want to study the effects of irradiation (e.g. alpha,
protons) on
> the level of individual cells. We are able to detect single particle
tracks
> with a track detector (basically a photographic emulsion) which is
> irradiated together with the cells or embryos with a vertical beam
(normal
> to the plane of the detector).
> With a microscope we take pictures of both the cells or embryos and
the
> developed detector and superimpose the two images. To tell which cell
> nuclei have been hit by a particle we must of course be able to see
the
> boundary of the nucleus (we would like to get away without
fluorescent
> staining).
> I have always had difficulties observing the nucleus in brightfield
or
> phase contrast (we don't have Nomarski and Hoffman is no solution) in
my
> B6C3F1 embryos. By chance I found out that in an other mouse strain
(NMRI)
> the nucleus is clearly visible even with bright field illumination.
> Unfortunatly the NMRI embryos didn't develop beyond the 4-cell stage
under
> my plain vanilla culture conditions which are OK for the B6C3F1.
>
> My question therefore: Is anybody aware of a mouse strain, where the
nuclei
> of life two-cell embryos are easily visible (unstained, no Nomarksi)
and
> which is healthy enough, so that the embryos will develop in vitro to
the
> blastocyst stage and hatch under standard culture conditions?
>
> Any help is highly appreciated.
B6C3F1 ova are somewhat pigmented and a bit more difficult to work
with than other strains. We've worked with a few different strains
that have readily visible nuclei at the 2-cell stage. For a hybrid,
B6SJL or B6D2 should work, also outbred Swiss (CD1, CF1, ICR, etc),
as well as a host of other strains including inbreds, such as B6 and
FVB.



From pinkert at cmed.bhs.uab.edu Mon Sep 9 14:01:18 1996
From: pinkert at cmed.bhs.uab.edu (Dr. C.A. Pinkert)
Date: Wed Aug 20 11:52:52 2003
Subject: (Fwd) Re: visibility of cell nuclei in tw-cell mouse embryos
Message-ID: <19960909180604693.AAA95@cap1>

> My question therefore: Is anybody aware of a mouse strain, where the
nuclei
> of life two-cell embryos are easily visible (unstained, no Nomarksi)
and
> which is healthy enough, so that the embryos will develop in vitro to
the
> blastocyst stage and hatch under standard culture conditions?
>
> Any help is highly appreciated.
B6C3F1 ova are somewhat pigmented and a bit more difficult to work
with than other strains. We've worked with a few different strains
that have readily visible nuclei at the 2-cell stage. For a hybrid,
B6SJL or B6D2 should work, also outbred Swiss (CD1, CF1, ICR, etc.),
as well as a host of other strains including inbreds, such as C57BL/6
and
FVB - although the viability of the B6 ova would not be equivalent to
the other strains.    C.A. Pinkert



From tjf at uci.edu Mon Sep 9 12:40:31 1996
From: tjf at uci.edu (Tom Fielder)
Date: Wed Aug 20 11:52:52 2003
Subject: fertilization of mouse eggs
Message-ID: <v03007804ae5a150547a8@[128.200.21.145]>

I am using FVB and B6D2F1 mice to make transgenics. I just started
superovulating the B6D2's last week, and I intended to mate them with
B6D2
males, but since the males weren't quite old enough, I decided to mate
them
with some proven FVB studs. I've done this twice, and both times I got
a
very low percentage of fertilized eggs. Both times there were obvious
plugs in 5/7 females, and the yield of eggs per oviduct varied quite a
bit
from mouse to mouse. The first time, 20% of the eggs were fertilized,
and
the second time only 10%, as judged by the presence of 2 pronuclei. Is
there some reason why the B6D2F1 X FVB mating would produce this
result?
The males are not more than 5 months old and have produced very high
percentages of fertilized eggs when mated with FVB females. All other
factors are equal (same lot of gonadotropins, same room, same feed,
same
light cycle, same timing of injections). Any ideas or comments would
be
greatly appreciated.
Tom




From dbowtell at petermac.unimelb.edu.au Wed Sep 11 10:02:38 1996
From: dbowtell at petermac.unimelb.edu.au (David Bowtell)
Date: Wed Aug 20 11:52:52 2003
Subject: Mating of chimeric mice
Message-ID: <v03007800ae5c308c90e1@[203.4.164.101]>
We have a dozen or so chimeric mice from a KO in W9.5 cells. There is
a
strong sex distortion and some of the mice are very strong. This line
has
been used very successfully in a collaborating laboratory, with a high
frequency of germline transmission. The first few mice have been mated
but
have not plugged C57BL/6J female over a period of about 4 weeks (these
chimeric males are now ca. 12wo). Clearly we have a way to go before
we
have a problem but does anyone have any comments on 'wringing' the most
out
of reluctant chimeras?

Thanks.

******* Please note new email address **********
David Bowtell
Principal Research Fellow
Research Division
Peter MacCallum Cancer Institute
Locked Bag 1 A'Beckett St,
Melbourne 3000

Lab 61-3-96561287
Office 61-3-96561296
Fax 61-3-96561411
email: dbowtell@petermac.unimelb.edu.au




From pinkert at cmed.bhs.uab.edu Tue Sep 10 19:18:00 1996
From: pinkert at cmed.bhs.uab.edu (Dr. C.A. Pinkert)
Date: Wed Aug 20 11:52:52 2003
Subject: Mating of chimeric mice
Message-ID: <19960910232244761.AAA62@cap1>

> have not plugged C57BL/6J female over a period of about 4 weeks
(these
> chimeric males are now ca. 12wo). Clearly we have a way to go before
we
> have a problem but does anyone have any comments on 'wringing' the
most out
> of reluctant chimeras?
Replace the females with new mice (preferably in proestrus) or
setting up for an IVF protocol (using survival surgery and removal of
the vas deferens and epididymis from one side only - thereby
maintaining the male for future breeding, and if there is a problem
with the first attempt - you still have another chance at rescuing
the line). The IVF scheme can also provide an index of relative
fertility based on sperm concentration, morphology, and motility.
Carl A. Pinkert
From TSAUNDER at hg-basic1mail.hg.med.umich.edu Wed Sep 11 09:34:42
1996
From: TSAUNDER at hg-basic1mail.hg.med.umich.edu (TSAUNDER)
Date: Wed Aug 20 11:52:52 2003
Subject: Mating of chimeric mice
Message-ID: <3001E7680B@HG-BASIC1MAIL.HG.MED.UMICH.EDU>

David,

Our appraoch to chimeras is to give them them many mating
opportunities.
To wit: rotate 2 young C57BL/6 females (6-8 weeks old) through the
male's cage every two weeks. This assumes that a male will have enough
sperm for one fertile mating per week. Removed females are housed in
pairs until it is determined whether they are gravid. By the time the
third pair of females is introduced, you should have weaned pups from
the first pair. Alternatively, you may wish to consider the IVF
approach
suggested by Carl Pinkert.

Thom Saunders, Ph.D.
Transgenic Animal Model Core
Biomedical Research Core Facilities
University of Michigan Medical School
email: tsaunder@umich.edu


Date:          Wed, 11 Sep 1996 09:02:38 +1000
To:            transgenic-list@ic.ac.uk
From:          David Bowtell <dbowtell@petermac.unimelb.edu.au>
Subject:       Mating of chimeric mice
Reply-to:      transgenic-list@ic.ac.uk

We have a dozen or so chimeric mice from a KO in W9.5 cells. There is
a
strong sex distortion and some of the mice are very strong. This line
has been used very successfully in a collaborating laboratory, with a
high frequency of germline transmission. The first few mice have been
mated but have not plugged C57BL/6J female over a period of about 4
weeks (these chimeric males are now ca. 12wo). Clearly we have a way
to
go before we have a problem but does anyone have any comments on
'wringing' the most out of reluctant chimeras?

Thanks.

******* Please note new email address **********
David Bowtell
Principal Research Fellow
Research Division
Peter MacCallum Cancer Institute
Locked Bag 1 A'Beckett St,
Melbourne 3000

Lab 61-3-96561287
Office 61-3-96561296
Fax 61-3-96561411
email: dbowtell@petermac.unimelb.edu.au




From jparkert at magnus.acs.ohio-state.edu Wed Sep 11 09:41:57 1996
From: jparkert at magnus.acs.ohio-state.edu (Janice Parker-Thornburg)
Date: Wed Aug 20 11:52:52 2003
Subject: Mating of chimeric mice
Message-ID: <199609111241.IAA07017@mail0.uts.ohio-state.edu>

>We have a dozen or so chimeric mice from a KO in W9.5 cells. There is
a
>strong sex distortion and some of the mice are very strong. This line
has
>been used very successfully in a collaborating laboratory, with a high
>frequency of germline transmission. The first few mice have been
mated but
>have not plugged C57BL/6J female over a period of about 4 weeks (these
>chimeric males are now ca. 12wo). Clearly we have a way to go before
we
>have a problem but does anyone have any comments on 'wringing' the
most out
>of reluctant chimeras?

>David Bowtell


David:

When we have "reluctant" males, our favorite trick is to give them one
or
two superovulated females, reasoning that: 1) we are sure that the
females
are in heat, and 2) the "quality" of their ovulatory phase might be
better
(of course, this is pure conjecture. . .).

Good luck.

Jan Parker-Thornburg

Jan Parker-Thornburg, Manager
Transgenic Animal Facility
The Ohio State University
jparkert@magnus.acs.ohio-state.edu




From carton at murray.fordham.edu Wed Sep 11 11:28:42 1996
From: carton at murray.fordham.edu (Jill Carton)
Date: Wed Aug 20 11:52:52 2003
Subject: knockout mouse vector
Message-ID: <v02130503ae5c7ad6e393@[169.132.99.85]>

Hi!
I was wondering if anyone knows of a convenient vector for sale which
could
be used to generate a knockout transgene. I was thinking of a vector
which
contained the HSV-TK gene, and a neomycin resistance gene surrounded by
two
unique multiple cloning sites for homologous sequence insertion.
Thanks.

Jill Carton
Dept. Biological Sciences
Fordham University
Larkin Hall, rm 160
Bronx, NY 10458
718-817-3652
718-817-3645 (fax)
carton@murray.fordham.edu




From mlm at titus.u-strasbg.fr Thu Sep 12 12:20:39 1996
From: mlm at titus.u-strasbg.fr (Marianne LEMEUR)
Date: Wed Aug 20 11:52:52 2003
Subject: NOD mice embryos
Message-ID: <v01510100ae5d9fea8c22@[130.79.78.57]>

Has anyone an idea how to improve yields of NOD embryos. Superovulation
is
impossible and natural matings seem to be productive only in a narrow
window.

Is there an hybrid cross which works well? We have tried B6XNOD which
is
not better.


Marianne Le Meur
Transgenic Animal Facility
IGBMC-Strasbourg, France




From pinkert at cmed.bhs.uab.edu Thu Sep 12 09:59:26 1996
From: pinkert at cmed.bhs.uab.edu (Dr. C.A. Pinkert)
Date: Wed Aug 20 11:52:52 2003
Subject: NOD zygote yields
Message-ID: <19960912140427737.AAA136@cap1>
> Has anyone an idea how to improve yields of NOD embryos.
Superovulation is
> impossible and natural matings seem to be productive only in a narrow
> window.
Marianne:
     As per Jackson Labs, superovulation actually works better in the
NOD's with older females (evidently 8-12 weeks is better than 3-4
weeks).
     In our first attempt this year (with approx. 8 week old females),
3
of 23 females were plugged and yielded 60 injectable eggs, of which
29 survived, and resulting in 1 stillborn and 2 resorbing fetuses.
     We held mice for 3 more weeks, and in our second attempt, 7 of 28
females (including some a second time) were plugged, yielding 145
injectable eggs, of which 120 survived. There were 19 liveborn pups,
18 weaned, and 4 founder transgenics.
     (Last year 163 eggs were injected on 2 separate days, 132
survived and were transferred, yielding 7/29 transgenics, but we went
through a goodly number of mice.)
     AI or IVF might be more beneficial, but the time to work out the
additional conditions/scheduling combined with our poor IVF yields
and variable timing with some BALB/c congenics and F1's dissuaded us
from going ahead in that direction.
     Good luck.     Carl



From TSAUNDER at hg-basic1mail.hg.med.umich.edu Fri Sep 13 08:31:13
1996
From: TSAUNDER at hg-basic1mail.hg.med.umich.edu (TSAUNDER)
Date: Wed Aug 20 11:52:52 2003
Subject: Forwarded: Ovulation Model
Message-ID: <5EF4E96C78@HG-BASIC1MAIL.HG.MED.UMICH.EDU>


This message posted on behalf of Jennifer Bowen,
Please direct replies to bowenjm@umich.edu
 -Thanks, Thom Saunders


I am interested in using a mouse model consisting of immature mice
stimulated to ovulate a normal or near-normal number of follicles (ie
not superovulated) and then mated to vasectomized males to induce
pseudopregnancy. I am designing this model for future use in a mutant
which is on a C57BL/6J X C3HB/FeJ background so will probably use
C57BL/6 animals. I would appreciate any information on what sort of
hormone dose/age combination will result in a normal number of
ovulations in immature females of this or similar strains.

Jennifer Bowen (bowenjm@umich.edu)




From mlm at titus.u-strasbg.fr Fri Sep 13 17:34:49 1996
From: mlm at titus.u-strasbg.fr (Marianne LEMEUR)
Date: Wed Aug 20 11:52:52 2003
Subject: sending frozen embryos
Message-ID: <v01510102ae5f3a5df9b5@[130.79.78.57]>

I would like to send frozen embryos all over the world instead of
sending
living mice, which is very time consuming to get all the formalities
together.

What are the conditions to do that, and were is it possible to buy the
container.
In France, our transportation company told us that the sender is
personnally responsible in case of an accident with liquid nitrogen
during
the flight!

Marianne Le Meur
IGBMC
Strasbourg-France




From ucympek at ucl.ac.uk Fri Sep 13 17:47:32 1996
From: ucympek at ucl.ac.uk (Peter Koder)
Date: Wed Aug 20 11:52:52 2003
Subject: sending frozen embryos
Message-ID: <E0v1aFi-0004AU-00@bowmore.cc.ic.ac.uk>

Hello Marianne

For a UK> recipient an Import licence is required from the Ministry of
Agriculture tel (0)181 330 8178 under the Importation of Embryos, Ova
and
Semen order. Also the permission of a Home Office Inspector under the
UK
Animals(Scientific Procedures) Act 1986

The importing institute Genetically Modified Organism (GMO) safety
committee
has also to be informed inorder to accept them under the Environmental
Protection Act (Contained Use) Regulations.

I hope this helps, Peter

I would like to send frozen embryos all over the world instead of
sending
>living mice, which is very time consuming to get all the formalities
>together.
>
>What are the conditions to do that, and were is it possible to buy the
>container.
>In France, our transportation company told us that the sender is
>personnally responsible in case of an accident with liquid nitrogen
during
>the flight!
>
>Marianne Le Meur
>IGBMC
>Strasbourg-France
>
>
>
>
>
>


Peter C Koder BVMS MSc MRCVS
UCL Biological Services         E-MAIL: UCYMPEK@UCL.AC.UK
University College London       TEL:+44(0)171 391 1309
Malet Place                     FAX:+44(0)171 380 7837
LONDON, WC1E 6BT
UK




From pinkert at cmed.bhs.uab.edu Fri Sep 13 11:55:51 1996
From: pinkert at cmed.bhs.uab.edu (Dr. C.A. Pinkert)
Date: Wed Aug 20 11:52:52 2003
Subject: sending frozen embryos
Message-ID: <19960913160057705.AAA159@cap1>

>   Date:        Fri, 13 Sep 1996 16:34:49 +0000
>   To:          transgenic-list@ic.ac.uk
>   From:        mlm@titus.u-strasbg.fr (Marianne LEMEUR)
>   Subject:     sending frozen embryos
>   Reply-to:    transgenic-list@ic.ac.uk

> I would like to send frozen embryos all over the world instead of
sending
> living mice, which is very time consuming to get all the formalities
> together.
>
> What are the conditions to do that, and were is it possible to buy
the
> container.
> In France, our transportation company told us that the sender is
> personnally responsible in case of an accident with liquid nitrogen
during
> the flight!
You may want to check with Dr. Glenn Monastersky regarding shipment
of ova and related considerations. He was using the liquid nitrogen
shipping containers that contained an absorbant material - it
maintained temperature, yet there was no LN2 sloshing around at all.
(what they used I believe was similar to the Arctic Express from
Thermolyne (model CY50915) that keeps temperature for 11 days) .
Glenn can be contacted at gmmgmm@aol.com.            Carl




From andreja at ariel.ucs.unimelb.edu.au   Sat Sep 14 11:09:28 1996
From: andreja at ariel.ucs.unimelb.edu.au (Andreja Pirkmaier)
Date: Wed Aug 20 11:52:52 2003
Subject: Caesarean sections
Message-ID: <199609140009.KAA10413@ariel.ucs.unimelb.EDU.AU>

       Has anyone any experience with injecting mice with hormones(
Day,
Dose) few Days prior doing caesarean section to avoid any early
delivery.
          I know that the gestation period for the transgenic line I am
interested in is 19 Days. I planed to do sections on Day 18 p. c..

Thanks Andreja Pirkmaier
Peter MacCallum Cancer Institute
andreja@ariel.ucs.unimelb.edu.au




From tonyjames at cuhk.edu.hk Sat Sep 14 10:43:18 1996
From: tonyjames at cuhk.edu.hk (A E James)
Date: Wed Aug 20 11:52:52 2003
Subject: Caesarean sections
Message-ID: <199609140143.JAA17319@hpg50a.csc.cuhk.edu.hk>

>        Has anyone any experience with injecting mice with hormones(
Day,
>Dose) few Days prior doing caesarean section to avoid any early
delivery.
>          I know that the gestation period for the transgenic line I
am
>interested in is 19 Days. I planed to do sections on Day 18 p. c..
>
>Thanks Andreja Pirkmaier
>Peter MacCallum Cancer Institute
>andreja@ariel.ucs.unimelb.edu.au
>
>
>
>Try Catherine O'Brien at WEHI or her senior technician at WEHI
Kew...her
name is Therese Johns.
Any way my protocol is progesterone for three days before the expected
birth. In pregnant rats the figure quoted is 20 mg per day for a heavy
preg.
rat.
Other figures I have seen for mice are 1.5 to 2.5 mg per day per mouse
for
the three days prior to csaerian delivery.
Hope this helps
Tony James
Tony James
BVSc MACVSc
Director
Laboratory Animal Unit
Chinese University of Hong Kong
Shatin, New Territories, Hong Kong
ph: 852 2609 6862
fx: 852 2603 5723
email: tonyjames@cuhk.edu.hk
.....................................
"nothing great was achieved without enthusiasm."
R. W. Emerson




From pinkert at cmed.bhs.uab.edu Sat Sep 14 09:48:18 1996
From: pinkert at cmed.bhs.uab.edu (Dr. C.A. Pinkert)
Date: Wed Aug 20 11:52:52 2003
Subject: Caesarean sections
Message-ID: <19960914135340470.AAA136@cap1>

> Has anyone any experience with injecting mice with hormones( Day,
> Dose) few Days prior doing caesarean section to avoid any early
delivery.
> I know that the gestation period for the transgenic line I am
> interested in is 19 Days. I planed to do sections on Day 18 p. c..
If the line uniformly delivers on d19 and the pups are robust, then
C-sections on d18 should likely work out well without the additional
hormone therapy.



From mlm at titus.u-strasbg.fr Tue Sep 17 16:19:26 1996
From: mlm at titus.u-strasbg.fr (Marianne LEMEUR)
Date: Wed Aug 20 11:52:52 2003
Subject: zygote yields
Message-ID: <v01510103ae646c5d8502@[130.79.78.57]>

Thanks to those who gave the solution for NOD mice zygote yield!
More generally it seems that C57Bl6 is the only inbred strain which can
be
superovulated in the same way as an F1 hybrid.
Other inbred strains like FVB/N or outbred strains like CD1 do not give
better yields than can be achieved with a natural mating.Is that
correct?
Marianne Le Meur




From jparkert at magnus.acs.ohio-state.edu Tue Sep 17 11:45:24 1996
From: jparkert at magnus.acs.ohio-state.edu (Janice Parker-Thornburg)
Date: Wed Aug 20 11:52:52 2003
Subject: zygote yields
Message-ID: <199609171445.KAA19688@postbox.acs.ohio-state.edu>

>Thanks to those who gave the solution for NOD mice zygote yield!
>More generally it seems that C57Bl6 is the only inbred strain which
can be
>superovulated in the same way as an F1 hybrid.
>Other inbred strains like FVB/N or outbred strains like CD1 do not
give
>better yields than can be achieved with a natural mating.Is that
correct?
>Marianne Le Meur

Marianne:

This is not correct. I have superovulated FVB/N for microinjection, as
well as for general harvesting of mouse embryos. If done in the
correct
time frame, one can get 8-10 more eggs (embryos) per female than with
natural matings. I also know that people using CD1's for embryo
harvesting
will superovulate them, presumably because they get a better yield. If
you
find that your yield does not improve with superovulation, it is likely
that your mouse-age or timespan for the hormone injections needs to be
optimized.

Jan Parker-Thornburg

Jan Parker-Thornburg, Manager
Transgenic Animal Facility
The Ohio State University
jparkert@magnus.acs.ohio-state.edu




From landel at medsci.udel.edu Tue Sep 17 12:37:11 1996
From: landel at medsci.udel.edu (Carlisle Landel)
Date: Wed Aug 20 11:52:53 2003
Subject: zygote yields
Message-ID: <199609171537.LAA03938@helios.medsci.udel.edu>

>Thanks to those who gave the solution for NOD mice zygote yield!
>More generally it seems that C57Bl6 is the only inbred strain which
can be
>superovulated in the same way as an F1 hybrid.
>Other inbred strains like FVB/N or outbred strains like CD1 do not
give
>better yields than can be achieved with a natural mating.Is that
correct?
>Marianne Le Meur

Marianne,

No, this isn't correct! I superovulate FVB/N routinely, and I'm pretty
sure that colleagues of mine in the past have done it to CD1's, too,
though
I vaguely recall there was some trick with dosage or timing or
something
(but I may be wrong--this was many years ago).

Check your hormones for proper concentration, and check that the light
controls in your facility are properly set, and make sure that you
are injecting at or very near the middle of your light cycle.

Regards,

Carlisle Landel
Dept. of Clinical Science
A.I. duPont Institute
PO Box 269
Wilmington DE 19899
(302) 651-6873
landel@helios.medsci.udel.edu
http://mdblmac.medsci.udel.edu/WEB/MDBL/Carlisle.html



From pinkert at cmed.bhs.uab.edu Tue Sep 17 10:23:08 1996
From: pinkert at cmed.bhs.uab.edu (Dr. C.A. Pinkert)
Date: Wed Aug 20 11:52:53 2003
Subject: zygote yields
Message-ID: <19960917142758010.AAB158@cap1>

> More generally it seems that C57Bl6 is the only inbred strain which
can be
> superovulated in the same way as an F1 hybrid.
I'm not sure what is meant here, but other inbred strains can be
utilized, using the same general scheme and sometimes jogging the
schedule a bit.

> Other inbred strains like FVB/N or outbred strains like CD1 do not
give
> better yields than can be achieved with a natural mating.Is that
correct?
It's a matter of degree, but I disagree with the last statement
(unless you mean that hybrids provide better net yields than either
inbreds and outbreds, which is true). In some labs, natural matings
facilitate a specific routine. However, there are significant
economies-of-scale when superovulation is employed (in relation to
zygote yields and needed labor/space/etc.) when using the strains
that you have mentioned.



From browng at medicine.wustl.edu Tue Sep 17 14:03:45 1996
From: browng at medicine.wustl.edu (Gary Brown)
Date: Wed Aug 20 11:52:53 2003
Subject: zygote yields
In-Reply-To: <v01510103ae646c5d8502@[130.79.78.57]>
Message-ID: <Pine.GSO.3.94.960917125610.29137A-100000@medicine>



On Tue, 17 Sep 1996, Marianne LEMEUR wrote:

> Thanks to those who gave the solution for NOD mice zygote yield!
> More generally it seems that C57Bl6 is the only inbred strain which
can be
> superovulated in the same way as an F1 hybrid.
> Other inbred strains like FVB/N or outbred strains like CD1 do not
give
> better yields than can be achieved with a natural mating.Is that
correct?
> Marianne Le Meur
>
>
>
>
Not so. I used FVB/N strain mice for 3 1/2 years and superovulated with
(in my opinion) a high degree of success. Since it's a larger mouse
than
B6C3F1 hybrids, we used 7.5IU hormones rather than 5IU, and a 47 hr gap
between PMSG and HCG. Typical yields were 25-30 good (non fragmented)
eggs per mouse. Can't say I've had experience with other strains other
than C57Bl/6 and B6C3F1 hybrids. Actually, I lie. 129SvJ mice are
dismal
for superovulation - if anyone can give me a way to do these
reproducibly
I'll get you a beer (or three)!

Gary Brown
E-mail: browng@medicine.wustl.edu or 75017.1050@compuserve.com
Web: http://ourworld.compuserve.com/homepages/TheBroons/
         "The Microinjection Workshop"




From carboni at valoritech.com Tue Sep 17 16:25:09 1996
From: carboni at valoritech.com (Nick Carboni)
Date: Wed Aug 20 11:52:53 2003
Subject: Wood's coculture technique
Message-ID: <199609171925.PAA25906@nash.pubnix.net>

Hi everybody,


Short message to know if one of you is currently working or has tried
to
work on wood's coculture technique.

I have spoke with some people, and it appears that it is rather
difficult to
acheive.

Has anyone any experience to share with me ??

Thanks in advance,

Nick
------------------------------------------------------------------
Nick Carboni, VP
Valoritech
1253, McGill College Av., room 620
Montreal, Quebec, Canada
H3B 2Y5
Tel.: (514) 866-8271 / Fax: (514) 866-8272
E-Mail: carboni@valoritech.com




From browng at medicine.wustl.edu Wed Sep 18 08:26:12 1996
From: browng at medicine.wustl.edu (Gary Brown)
Date: Wed Aug 20 11:52:53 2003
Subject: Wood's coculture technique
In-Reply-To: <199609171925.PAA25906@nash.pubnix.net>
Message-ID: <Pine.GSO.3.94.960918070739.1409A-100000@medicine>



On Tue, 17 Sep 1996, Nick Carboni wrote:

> Hi everybody,
>
>
> Short message to know if one of you is currently working or has tried
to
> work on wood's coculture technique.
>
> I have spoke with some people, and it appears that it is rather
difficult to
> acheive.
>
> Has anyone any experience to share with me ??
>
> Thanks in advance,
>
> Nick
>
Hi !

I tried Wood's co-culture technique around 3 years ago, but only had 1
recorded birth in 16 implanted mothers, and this pup was malformed. We
were unable to determine the efficacy of the technique as the ES cell
line
that we used was obtained from another laboratory, and both their
technique and the cell line itself were called into question. However,
here's what I can tell you..

1. We found that the ES cells did not adhere to the de - ZP'd morulae /
octoploid / tetraploid embryos as well as we thought the paper
indicated.

2. Our understanding is that the technique's success is very ES cell
dependent.

3. Our understanding is that the live birth rate is very low, making
this
a system better suited to in utero developmental studies.
Since this time, I have tried to find the time to explore non-injection
methods of chimera production (with little success on the time front!).
I
have only managed to complete a pilot study of Nagy / Rossant's
"sandwich"
method where I was successful in fusing 2 morulae together in culture
overnight in custom made dimples. This effort would have been more
instructive if I had had access to some ES cells to "sandwich", but the
fusions had a >90% success rate. For the reference on this
methodology,
do a Medline search on the authors above or check out the E-newsletter
archive on my homepage for August / September.

Hope this is instructive!     :-)

Gary Brown
E-mail: browng@medicine.wustl.edu or 75017.1050@compuserve.com
Web: http://ourworld.compuserve.com/homepages/TheBroons/
            "The Microinjection Workshop"




From ldeg at midway.uchicago.edu Wed Sep 18 21:28:26 1996
From: ldeg at midway.uchicago.edu (Linda Degenstein)
Date: Wed Aug 20 11:52:53 2003
Subject: ES Cells from Balb C mice
Message-ID: <199609190128.UAA22947@midway.uchicago.edu>

   We are looking for ES Cells (germline proven) that are made from
Balb C
strain mice. Does anyone know if these are available?

Linda Degenstein
Director UCCRC
Transgenic/ES Cell Facility
The University of Chicago
ldeg@midway.uchicago.edu




From khelmuth at facstaff.wisc.edu Tue Sep 24 15:28:52 1996
From: khelmuth at facstaff.wisc.edu (khelmuth@facstaff.wisc.edu)
Date: Wed Aug 20 11:52:53 2003
Subject: noise problems
Message-ID: <v02140b01ae6d9fec8a48@[144.92.94.130]>

Hi everyone:

I am looking for documentation that excess noise can affect the
breeding
and production of embryos from mice. Does anyone know of published
work
that looked at environmental affects on a mouse colony? Any citations
would be appreciated.
Kathy

Kathy Krentz Helmuth
Transgenic Facility
425 Henry Mall
Room 2210 Biotechnology Center
Madison, WI 53706
(608) 265-2801
email:khelmuth@facstaff.wisc.edu




From mlp at informatics.jax.org Tue Sep 24 17:20:46 1996
From: mlp at informatics.jax.org (Moyha Lennon-Pierce)
Date: Wed Aug 20 11:52:53 2003
Subject: Noise/reproduction
Message-ID: <v01540b05ae6dfd6df569@[192.233.41.18]>

The following references might be useful.

I searched the Strain Characteristics Catalog in development at The
Jackson
Lab and did a quick search in NLM. Not much at all in the literature,
but
the following references may be useful.

Nawrot PS et al.
Embryotoxicity of various noise stimuli in the mouse.
Teratology 1980; 22(3):279-89.
CF-1

Zakem HB, Alliston CW.
The effects of noise level and elevated ambient temperatures upon
selected
reproductive traits in female Swiss-Webster mice.
Lab Anim Sci 1974; 24:469-75.
Strain SWR

Moyha Lennon-Pierce
MGD Informatics
mlp@informatics.jax.org

>Hi everyone:
>
>I am looking for documentation that excess noise can affect the
breeding
>and production of embryos from mice. Does anyone know of published
work
>that looked at environmental affects on a mouse colony? Any citations
>would be appreciated.
>
>Kathy
>
>Kathy Krentz Helmuth
>Transgenic Facility
>425 Henry Mall
>Room 2210 Biotechnology Center
>Madison, WI 53706
>(608) 265-2801
>email:khelmuth@facstaff.wisc.edu




From landel at medsci.udel.edu Tue Sep 24 18:13:36 1996
From: landel at medsci.udel.edu (Carlisle Landel)
Date: Wed Aug 20 11:52:53 2003
Subject: Noise/reproduction
Message-ID: <199609242113.RAA08916@helios.medsci.udel.edu>

Kathy,

I saw a poster at a meeting a couple of weeks ago where they were using
one of those ultrasonic mouse repellers to increase the percentage
of embryo loss in CBAxDBA/2 model. If you want more info, I'll see if
I can point you towards the authors.

Regards,

Carlisle Landel
Dept. of Clinical Science
duPont Hospital for Children
PO Box 269
Wilmington DE 19899
(302) 651-6873
landel@helios.medsci.udel.edu
http://mdblmac.medsci.udel.edu/WEB/MDBL/Carlisle.html



From Roger.Leemann at imr.psi.ch Wed Sep 25 17:40:47 1996
From: Roger.Leemann at imr.psi.ch (Roger C. Leemann)
Date: Wed Aug 20 11:52:53 2003
Subject: noise problems
Message-ID: <324941f920c1002@pss200.psi.ch>

Kathy Krentz Helmuth (khelmuth@facstaff.wisc.edu) wrote:

>I am looking for documentation that excess noise can affect the
breeding
>and production of embryos from mice.

Kathy

Sorry, no literature, but I can tell you how we do it. In our animal
housing facility there is always a radio playing at a low volume during
the
daylight cycle (maybe this can not be referred to as "excessive"
noise.) I
don't know whether or how this affects the mice, positive or negative.
With
non-superovulated B6C3F1 mice I can expect 15% to 25% plugs and 7 to 12
two-cell embryos/mouse. How does this compare to other labs?

Roger

:::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::
:::::
Work: Institute for Medical Radiobiology        ++41 (56) 310-3792
[desk]
       CH-5232 Villigen PSI                     ++41 (1) 3856-561
[desk ZH]
       Switzerland                              ++41 (56) 310 3294
[fax]
Home: Nordstrasse 26, CH-8006 Zuerich
       Switzerland                              ++41 (1) 361 0349
[home]




From DAMAK at whio.lincoln.ac.nz Mon Sep 2 02:32:54 1996
From: DAMAK at whio.lincoln.ac.nz (Damak, Sami)

Subject: Import of transgenic mice into the US
Message-ID: <17CA2DC2C3E@whio.lincoln.ac.nz>

I am looking at sending transgenic mice from New Zealand to the US.
Does anyone know who I should approach to get customs and quarantine
clearance?
Thanks
Sami Damak



From Dnxrh at aol.com Tue Sep 3 04:07:29 1996
From: Dnxrh at aol.com (Dnxrh@aol.com)

Subject: Import of transgenic mice into the US (DNA Too!)
Message-ID: <960902230729_275590968@emout12.mail.aol.com>

If permits are required (they may not be) it is the responsibility of
the
importer (your US recipient) to get the permits. Have them contact the
USDA -
APHIS to determine if permits are needed.

USDA - APHIS can be found at:
http://www.aphis.usda.gov
301-734-7885 (phone)
301-734-8226

The guidelines seem to change but at the present time you should know
the
following:
Do the mice contain any livestock or avian genes ?

Do the mice contain and genes from any livestock or avian pathogens?

Are the mice infected with any known diseases which are transferable to
humans, birds or livestock?

Do the mice contain any genes from human pathogens?

Have the mice been treated with any human blood products or any
livestock
derived products (ie. albumin).

If you can answer "no" to these questions then your recipient in the US
will
probably will not need a permit. Even if a permit is not required, the
animals will still need to have paperwork documenting where they come
from,
what genes they contain and what their current health status is.

If you do need a permit they can take 6 weeks to get.

I have had more experience shipping mice out of the US than in shipping
them
into the US. I suggest very strongly that you or your recipient in the
US
contact USDA-APHIS directly to confirm this information.

PS. to all you gene jockeys out there. These same rules apply to all
recombinant DNA as well. We have had plasmids and DNA fragments held up
by
customs because the invoices did not state clearly all of the above.

Good Luck,
Rick Huntress
DNX Transgenics
dnxrh@aol.com
+508-779-0189 (phone)
+508-779-0190 (fax)

My opinions are my own and not necessarily that of my employer.




From WRIGHTA at gunet.georgetown.edu Tue Sep 3 17:07:11 1996
From: WRIGHTA at gunet.georgetown.edu (Ann Wright)

Subject: Import of transgenic mice into the US -Reply
Message-ID: <s22c13c8.006@gunet.georgetown.edu>

Information regarding APHIS programs and import-export permits can be
obtained on the home page http://www.aphis.usda.gov. The fax number
is 301-734-8226, email ncie@aphis.usda.gov or voice mail
301-734-4412. The permit to import animals is needed to clear customs.
From carton at murray.fordham.edu Wed Sep 4 00:02:12 1996
From: carton at murray.fordham.edu (Jill Carton)

Subject: electroporators for ES cells
Message-ID: <v02130500ae5263ad7834@[169.132.99.88]>

I am a graduate student at Fordham University working on generating a
knockout mouse. The electroporation protocols I have found for the ES
cells are all using the Biorad Gene Pulser. We have in our department
a
Baekon 2000 gene transfer system. I was wondering if anyone has ever
used
this apparatus for electroporation of ES cells and if you would share
your
protocol with me. This would save me a lot of time trying to work out
the
conditions.

Thanks in advance for the information,
              Jill Carton

Jill Carton
Dept. Biological Sciences
Fordham University
Larkin Hall, rm 160
Bronx, NY 10458
718-817-3652
718-817-3645 (fax)
carton@murray.fordham.edu




From jklohse at facstaff.wisc.edu Fri Sep 6 23:57:44 1996
From: jklohse at facstaff.wisc.edu (Jan K. Lohse)

Subject: transgene control systems
Message-ID: <v02140b01ae565caab1f7@[144.92.48.83]>


I'm interested in finding out what sorts of transgenic mice people are
currently creating with non-leaky, induceable promoter systems such as
the
ones that use the tet operon. Are these being used with good success?
Satisfactory expression, turned on or off by a drug as designed? Is
the
lac operon being used for this sort of thing as well?   Any
descriptions of
constructs, suggestions, and info about potential problems you're
willing
to share would be appreciated.
Also, how about similar info about recombination modification systems
such
as CRE-LOX and FLP?

Any other cool control systems out there?

Thanks very much.   I think many of us will find the answers
interesting.

Jan




From fmargoli at umabnet.ab.umd.edu Mon Sep 9 15:36:33 1996
From: fmargoli at umabnet.ab.umd.edu (Frank L. Margolis)

Subject: transgene control systems
Message-ID: <v01530500ae59db81c04d@[134.192.49.38]>

>I'm interested in finding out what sorts of transgenic mice people are
>currently creating with non-leaky, induceable promoter systems such as
the
>ones that use the tet operon. Are these being used with good success?
>Satisfactory expression, turned on or off by a drug as designed? Is
the
>lac operon being used for this sort of thing as well?   Any
descriptions of
>constructs, suggestions, and info about potential problems you're
willing
>to share would be appreciated.
>
>Also, how about similar info about recombination modification systems
such
>as CRE-LOX and FLP?
>
>Any other cool control systems out there?
>
>Thanks very much. I think many of us will find the answers
interesting.
>
>Jan
Similar questions arise with regard to the ecdysone regulated promoter
recently reported in PNAS. Any info would be useful to all.

Frank Margolis

Frank L. Margolis Ph.D.
Department of Anatomy
University of Maryland at Baltimore
School of Medicine
685 West Baltimore Street
Baltimore MD 21201

Phone 410-706-8913 office
             -8914 lab
FAX          -2512 Department office

"There is something fascinating about science: one gets such wholesale
returns of conjecture out of such a trifling investment of fact."
Mark
Twain




From Roger.Leemann at imr.psi.ch Mon Sep 9 18:37:54 1996
From: Roger.Leemann at imr.psi.ch (Roger C. Leemann)

Subject: visibility of cell nuclei in tw-cell mouse embryos
Message-ID: <323453844d7e002@pss200.psi.ch>

Hi

Gary Brown came across a few questions I had sent to the rodent-
research
list and because he had to admit that his answers to most of them would
be
"I don't know" suggested I might give it a try on this list.

Sure, I'd like to, although it's only indirectly related to transgenic
mice. Thanks Gary.

I use 2-cell embryos of B6C3F1xB6C3F1 (Charles River, Germany) in a
project
where we want to study the effects of irradiation (e.g. alpha, protons)
on
the level of individual cells. We are able to detect single particle
tracks
with a track detector (basically a photographic emulsion) which is
irradiated together with the cells or embryos with a vertical beam
(normal
to the plane of the detector).
With a microscope we take pictures of both the cells or embryos and the
developed detector and superimpose the two images. To tell which cell
nuclei have been hit by a particle we must of course be able to see the
boundary of the nucleus (we would like to get away without fluorescent
staining).
I have always had difficulties observing the nucleus in brightfield or
phase contrast (we don't have Nomarski and Hoffman is no solution) in
my
B6C3F1 embryos. By chance I found out that in an other mouse strain
(NMRI)
the nucleus is clearly visible even with bright field illumination.
Unfortunatly the NMRI embryos didn't develop beyond the 4-cell stage
under
my plain vanilla culture conditions which are OK for the B6C3F1.

My question therefore: Is anybody aware of a mouse strain, where the
nuclei
of life two-cell embryos are easily visible (unstained, no Nomarksi)
and
which is healthy enough, so that the embryos will develop in vitro to
the
blastocyst stage and hatch under standard culture conditions?

Any help is highly appreciated.

Roger

:::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::
:::::
Work: Institute for Medical Radiobiology        ++41 (56) 310-3792
[desk]
       CH-5232 Villigen PSI                     ++41 (1) 3856-561
[desk ZH]
       Switzerland                              ++41 (56) 310 3294
[fax]
Home: Nordstrasse 26, CH-8006 Zuerich
       Switzerland                              ++41 (1) 361 0349
[home]




From pinkert at cmed.bhs.uab.edu Mon Sep 9 13:56:56 1996
From: pinkert at cmed.bhs.uab.edu (Dr. C.A. Pinkert)

Subject: visibility of cell nuclei in tw-cell mouse embryos
Message-ID: <19960909180142886.AAA77@cap1>

>   Date:        Mon, 9 Sep 1996 19:37:54 +0200
>   To:          transgenic-list@ic.ac.uk
>   From:        Roger.Leemann@imr.psi.ch (Roger C. Leemann)
>   Subject:     visibility of cell nuclei in tw-cell mouse embryos
>   Reply-to:    transgenic-list@ic.ac.uk

> Hi
>
> Gary Brown came across a few questions I had sent to the rodent-
research
> list and because he had to admit that his answers to most of them
would be
> "I don't know" suggested I might give it a try on this list.
>
> Sure, I'd like to, although it's only indirectly related to
transgenic
> mice. Thanks Gary.
>
> I use 2-cell embryos of B6C3F1xB6C3F1 (Charles River, Germany) in a
project
> where we want to study the effects of irradiation (e.g. alpha,
protons) on
> the level of individual cells. We are able to detect single particle
tracks
> with a track detector (basically a photographic emulsion) which is
> irradiated together with the cells or embryos with a vertical beam
(normal
> to the plane of the detector).
> With a microscope we take pictures of both the cells or embryos and
the
> developed detector and superimpose the two images. To tell which cell
> nuclei have been hit by a particle we must of course be able to see
the
> boundary of the nucleus (we would like to get away without
fluorescent
> staining).
> I have always had difficulties observing the nucleus in brightfield
or
> phase contrast (we don't have Nomarski and Hoffman is no solution) in
my
> B6C3F1 embryos. By chance I found out that in an other mouse strain
(NMRI)
> the nucleus is clearly visible even with bright field illumination.
> Unfortunatly the NMRI embryos didn't develop beyond the 4-cell stage
under
> my plain vanilla culture conditions which are OK for the B6C3F1.
>
> My question therefore: Is anybody aware of a mouse strain, where the
nuclei
> of life two-cell embryos are easily visible (unstained, no Nomarksi)
and
> which is healthy enough, so that the embryos will develop in vitro to
the
> blastocyst stage and hatch under standard culture conditions?
>
> Any help is highly appreciated.
B6C3F1 ova are somewhat pigmented and a bit more difficult to work
with than other strains. We've worked with a few different strains
that have readily visible nuclei at the 2-cell stage. For a hybrid,
B6SJL or B6D2 should work, also outbred Swiss (CD1, CF1, ICR, etc),
as well as a host of other strains including inbreds, such as B6 and
FVB.



From pinkert at cmed.bhs.uab.edu Mon Sep 9 14:01:18 1996
From: pinkert at cmed.bhs.uab.edu (Dr. C.A. Pinkert)

Subject: (Fwd) Re: visibility of cell nuclei in tw-cell mouse embryos
Message-ID: <19960909180604693.AAA95@cap1>

> My question therefore: Is anybody aware of a mouse strain, where the
nuclei
> of life two-cell embryos are easily visible (unstained, no Nomarksi)
and
> which is healthy enough, so that the embryos will develop in vitro to
the
> blastocyst stage and hatch under standard culture conditions?
>
> Any help is highly appreciated.
B6C3F1 ova are somewhat pigmented and a bit more difficult to work
with than other strains. We've worked with a few different strains
that have readily visible nuclei at the 2-cell stage. For a hybrid,
B6SJL or B6D2 should work, also outbred Swiss (CD1, CF1, ICR, etc.),
as well as a host of other strains including inbreds, such as C57BL/6
and
FVB - although the viability of the B6 ova would not be equivalent to
the other strains.    C.A. Pinkert



From tjf at uci.edu Mon Sep 9 19:40:31 1996
From: tjf at uci.edu (Tom Fielder)

Subject: fertilization of mouse eggs
Message-ID: <v03007804ae5a150547a8@[128.200.21.145]>

I am using FVB and B6D2F1 mice to make transgenics. I just started
superovulating the B6D2's last week, and I intended to mate them with
B6D2
males, but since the males weren't quite old enough, I decided to mate
them
with some proven FVB studs. I've done this twice, and both times I got
a
very low percentage of fertilized eggs. Both times there were obvious
plugs in 5/7 females, and the yield of eggs per oviduct varied quite a
bit
from mouse to mouse. The first time, 20% of the eggs were fertilized,
and
the second time only 10%, as judged by the presence of 2 pronuclei. Is
there some reason why the B6D2F1 X FVB mating would produce this
result?
The males are not more than 5 months old and have produced very high
percentages of fertilized eggs when mated with FVB females. All other
factors are equal (same lot of gonadotropins, same room, same feed,
same
light cycle, same timing of injections). Any ideas or comments would
be
greatly appreciated.
Tom




From dbowtell at petermac.unimelb.edu.au Wed Sep 11 00:02:38 1996
From: dbowtell at petermac.unimelb.edu.au (David Bowtell)

Subject: Mating of chimeric mice
Message-ID: <v03007800ae5c308c90e1@[203.4.164.101]>

We have a dozen or so chimeric mice from a KO in W9.5 cells. There is
a
strong sex distortion and some of the mice are very strong. This line
has
been used very successfully in a collaborating laboratory, with a high
frequency of germline transmission. The first few mice have been mated
but
have not plugged C57BL/6J female over a period of about 4 weeks (these
chimeric males are now ca. 12wo). Clearly we have a way to go before
we
have a problem but does anyone have any comments on 'wringing' the most
out
of reluctant chimeras?

Thanks.

******* Please note new email address **********
David Bowtell
Principal Research Fellow
Research Division
Peter MacCallum Cancer Institute
Locked Bag 1 A'Beckett St,
Melbourne 3000

Lab 61-3-96561287
Office 61-3-96561296
Fax 61-3-96561411
email: dbowtell@petermac.unimelb.edu.au




From pinkert at cmed.bhs.uab.edu Tue Sep 10 19:18:00 1996
From: pinkert at cmed.bhs.uab.edu (Dr. C.A. Pinkert)

Subject: Mating of chimeric mice
Message-ID: <19960910232244761.AAA62@cap1>

> have not plugged C57BL/6J female over a period of about 4 weeks
(these
> chimeric males are now ca. 12wo). Clearly we have a way to go before
we
> have a problem but does anyone have any comments on 'wringing' the
most out
> of reluctant chimeras?
Replace the females with new mice (preferably in proestrus) or
setting up for an IVF protocol (using survival surgery and removal of
the vas deferens and epididymis from one side only - thereby
maintaining the male for future breeding, and if there is a problem
with the first attempt - you still have another chance at rescuing
the line). The IVF scheme can also provide an index of relative
fertility based on sperm concentration, morphology, and motility.
Carl A. Pinkert



From TSAUNDER at hg-basic1mail.hg.med.umich.edu Wed Sep 11 14:34:42
1996
From: TSAUNDER at hg-basic1mail.hg.med.umich.edu (TSAUNDER)

Subject: Mating of chimeric mice
Message-ID: <3001E7680B@HG-BASIC1MAIL.HG.MED.UMICH.EDU>

David,
Our appraoch to chimeras is to give them them many mating
opportunities.
To wit: rotate 2 young C57BL/6 females (6-8 weeks old) through the
male's cage every two weeks. This assumes that a male will have enough
sperm for one fertile mating per week. Removed females are housed in
pairs until it is determined whether they are gravid. By the time the
third pair of females is introduced, you should have weaned pups from
the first pair. Alternatively, you may wish to consider the IVF
approach
suggested by Carl Pinkert.

Thom Saunders, Ph.D.
Transgenic Animal Model Core
Biomedical Research Core Facilities
University of Michigan Medical School
email: tsaunder@umich.edu


Date:          Wed, 11 Sep 1996 09:02:38 +1000
To:            transgenic-list@ic.ac.uk
From:          David Bowtell <dbowtell@petermac.unimelb.edu.au>
Subject:       Mating of chimeric mice
Reply-to:      transgenic-list@ic.ac.uk

We have a dozen or so chimeric mice from a KO in W9.5 cells. There is
a
strong sex distortion and some of the mice are very strong. This line
has been used very successfully in a collaborating laboratory, with a
high frequency of germline transmission. The first few mice have been
mated but have not plugged C57BL/6J female over a period of about 4
weeks (these chimeric males are now ca. 12wo). Clearly we have a way
to
go before we have a problem but does anyone have any comments on
'wringing' the most out of reluctant chimeras?

Thanks.

******* Please note new email address **********
David Bowtell
Principal Research Fellow
Research Division
Peter MacCallum Cancer Institute
Locked Bag 1 A'Beckett St,
Melbourne 3000

Lab 61-3-96561287
Office 61-3-96561296
Fax 61-3-96561411
email: dbowtell@petermac.unimelb.edu.au
From jparkert at magnus.acs.ohio-state.edu Wed Sep 11 13:41:57 1996
From: jparkert at magnus.acs.ohio-state.edu (Janice Parker-Thornburg)

Subject: Mating of chimeric mice
Message-ID: <199609111241.IAA07017@mail0.uts.ohio-state.edu>

>We have a dozen or so chimeric mice from a KO in W9.5 cells. There is
a
>strong sex distortion and some of the mice are very strong. This line
has
>been used very successfully in a collaborating laboratory, with a high
>frequency of germline transmission. The first few mice have been
mated but
>have not plugged C57BL/6J female over a period of about 4 weeks (these
>chimeric males are now ca. 12wo). Clearly we have a way to go before
we
>have a problem but does anyone have any comments on 'wringing' the
most out
>of reluctant chimeras?

>David Bowtell


David:

When we have "reluctant" males, our favorite trick is to give them one
or
two superovulated females, reasoning that: 1) we are sure that the
females
are in heat, and 2) the "quality" of their ovulatory phase might be
better
(of course, this is pure conjecture. . .).

Good luck.

Jan Parker-Thornburg

Jan Parker-Thornburg, Manager
Transgenic Animal Facility
The Ohio State University
jparkert@magnus.acs.ohio-state.edu




From carton at murray.fordham.edu Wed Sep 11 15:28:42 1996
From: carton at murray.fordham.edu (Jill Carton)

Subject: knockout mouse vector
Message-ID: <v02130503ae5c7ad6e393@[169.132.99.85]>

Hi!
I was wondering if anyone knows of a convenient vector for sale which
could
be used to generate a knockout transgene. I was thinking of a vector
which
contained the HSV-TK gene, and a neomycin resistance gene surrounded by
two
unique multiple cloning sites for homologous sequence insertion.
Thanks.

Jill Carton
Dept. Biological Sciences
Fordham University
Larkin Hall, rm 160
Bronx, NY 10458
718-817-3652
718-817-3645 (fax)
carton@murray.fordham.edu




From mlm at titus.u-strasbg.fr Thu Sep 12 12:20:39 1996
From: mlm at titus.u-strasbg.fr (Marianne LEMEUR)

Subject: NOD mice embryos
Message-ID: <v01510100ae5d9fea8c22@[130.79.78.57]>

Has anyone an idea how to improve yields of NOD embryos. Superovulation
is
impossible and natural matings seem to be productive only in a narrow
window.

Is there an hybrid cross which works well? We have tried B6XNOD which
is
not better.


Marianne Le Meur
Transgenic Animal Facility
IGBMC-Strasbourg, France




From pinkert at cmed.bhs.uab.edu Thu Sep 12 09:59:26 1996
From: pinkert at cmed.bhs.uab.edu (Dr. C.A. Pinkert)

Subject: NOD zygote yields
Message-ID: <19960912140427737.AAA136@cap1>

> Has anyone an idea how to improve yields of NOD embryos.
Superovulation is
> impossible and natural matings seem to be productive only in a narrow
> window.
Marianne:
     As per Jackson Labs, superovulation actually works better in the
NOD's with older females (evidently 8-12 weeks is better than 3-4
weeks).
     In our first attempt this year (with approx. 8 week old females),
3
of 23 females were plugged and yielded 60 injectable eggs, of which
29 survived, and resulting in 1 stillborn and 2 resorbing fetuses.
     We held mice for 3 more weeks, and in our second attempt, 7 of 28
females (including some a second time) were plugged, yielding 145
injectable eggs, of which 120 survived. There were 19 liveborn pups,
18 weaned, and 4 founder transgenics.
     (Last year 163 eggs were injected on 2 separate days, 132
survived and were transferred, yielding 7/29 transgenics, but we went
through a goodly number of mice.)
     AI or IVF might be more beneficial, but the time to work out the
additional conditions/scheduling combined with our poor IVF yields
and variable timing with some BALB/c congenics and F1's dissuaded us
from going ahead in that direction.
     Good luck.     Carl



From TSAUNDER at hg-basic1mail.hg.med.umich.edu Fri Sep 13 13:31:13
1996
From: TSAUNDER at hg-basic1mail.hg.med.umich.edu (TSAUNDER)

Subject: Forwarded: Ovulation Model
Message-ID: <5EF4E96C78@HG-BASIC1MAIL.HG.MED.UMICH.EDU>


This message posted on behalf of Jennifer Bowen,
Please direct replies to bowenjm@umich.edu
 -Thanks, Thom Saunders


I am interested in using a mouse model consisting of immature mice
stimulated to ovulate a normal or near-normal number of follicles (ie
not superovulated) and then mated to vasectomized males to induce
pseudopregnancy. I am designing this model for future use in a mutant
which is on a C57BL/6J X C3HB/FeJ background so will probably use
C57BL/6 animals. I would appreciate any information on what sort of
hormone dose/age combination will result in a normal number of
ovulations in immature females of this or similar strains.

Jennifer Bowen (bowenjm@umich.edu)




From mlm at titus.u-strasbg.fr Fri Sep 13 17:34:49 1996
From: mlm at titus.u-strasbg.fr (Marianne LEMEUR)

Subject: sending frozen embryos
Message-ID: <v01510102ae5f3a5df9b5@[130.79.78.57]>

I would like to send frozen embryos all over the world instead of
sending
living mice, which is very time consuming to get all the formalities
together.
What are the conditions to do that, and were is it possible to buy the
container.
In France, our transportation company told us that the sender is
personnally responsible in case of an accident with liquid nitrogen
during
the flight!

Marianne Le Meur
IGBMC
Strasbourg-France




From ucympek at ucl.ac.uk Fri Sep 13 16:47:32 1996
From: ucympek at ucl.ac.uk (Peter Koder)

Subject: sending frozen embryos
Message-ID: <E0v1aFi-0004AU-00@bowmore.cc.ic.ac.uk>

Hello Marianne

For a UK> recipient an Import licence is required from the Ministry of
Agriculture tel (0)181 330 8178 under the Importation of Embryos, Ova
and
Semen order. Also the permission of a Home Office Inspector under the
UK
Animals(Scientific Procedures) Act 1986

The importing institute Genetically Modified Organism (GMO) safety
committee
has also to be informed inorder to accept them under the Environmental
Protection Act (Contained Use) Regulations.

I hope this helps, Peter

I would like to send frozen embryos all over the world instead of
sending
>living mice, which is very time consuming to get all the formalities
>together.
>
>What are the conditions to do that, and were is it possible to buy the
>container.
>In France, our transportation company told us that the sender is
>personnally responsible in case of an accident with liquid nitrogen
during
>the flight!
>
>Marianne Le Meur
>IGBMC
>Strasbourg-France
>
>
>
>
>
>


Peter C Koder BVMS MSc MRCVS
UCL Biological Services           E-MAIL: UCYMPEK@UCL.AC.UK
University College London         TEL:+44(0)171 391 1309
Malet Place                       FAX:+44(0)171 380 7837
LONDON, WC1E 6BT
UK




From pinkert at cmed.bhs.uab.edu Fri Sep 13 11:55:51 1996
From: pinkert at cmed.bhs.uab.edu (Dr. C.A. Pinkert)

Subject: sending frozen embryos
Message-ID: <19960913160057705.AAA159@cap1>

>   Date:          Fri, 13 Sep 1996 16:34:49 +0000
>   To:            transgenic-list@ic.ac.uk
>   From:          mlm@titus.u-strasbg.fr (Marianne LEMEUR)
>   Subject:       sending frozen embryos
>   Reply-to:      transgenic-list@ic.ac.uk

> I would like to send frozen embryos all over the world instead of
sending
> living mice, which is very time consuming to get all the formalities
> together.
>
> What are the conditions to do that, and were is it possible to buy
the
> container.
> In France, our transportation company told us that the sender is
> personnally responsible in case of an accident with liquid nitrogen
during
> the flight!
You may want to check with Dr. Glenn Monastersky regarding shipment
of ova and related considerations. He was using the liquid nitrogen
shipping containers that contained an absorbant material - it
maintained temperature, yet there was no LN2 sloshing around at all.
(what they used I believe was similar to the Arctic Express from
Thermolyne (model CY50915) that keeps temperature for 11 days) .
Glenn can be contacted at gmmgmm@aol.com.            Carl




From andreja at ariel.ucs.unimelb.edu.au Sat Sep 14 01:09:28 1996
From: andreja at ariel.ucs.unimelb.edu.au (Andreja Pirkmaier)

Subject: Caesarean sections
Message-ID: <199609140009.KAA10413@ariel.ucs.unimelb.EDU.AU>

         Has anyone any experience with injecting mice with hormones(
Day,
Dose) few Days prior doing caesarean section to avoid any early
delivery.
          I know that the gestation period for the transgenic line I am
interested in is 19 Days. I planed to do sections on Day 18 p. c..

Thanks Andreja Pirkmaier
Peter MacCallum Cancer Institute
andreja@ariel.ucs.unimelb.edu.au




From tonyjames at cuhk.edu.hk Sat Sep 14 02:43:18 1996
From: tonyjames at cuhk.edu.hk (A E James)

Subject: Caesarean sections
Message-ID: <199609140143.JAA17319@hpg50a.csc.cuhk.edu.hk>

>        Has anyone any experience with injecting mice with hormones(
Day,
>Dose) few Days prior doing caesarean section to avoid any early
delivery.
>          I know that the gestation period for the transgenic line I
am
>interested in is 19 Days. I planed to do sections on Day 18 p. c..
>
>Thanks Andreja Pirkmaier
>Peter MacCallum Cancer Institute
>andreja@ariel.ucs.unimelb.edu.au
>
>
>
>Try Catherine O'Brien at WEHI or her senior technician at WEHI
Kew...her
name is Therese Johns.
Any way my protocol is progesterone for three days before the expected
birth. In pregnant rats the figure quoted is 20 mg per day for a heavy
preg.
rat.
Other figures I have seen for mice are 1.5 to 2.5 mg per day per mouse
for
the three days prior to csaerian delivery.
Hope this helps
Tony James
Tony James
BVSc MACVSc
Director
Laboratory Animal Unit
Chinese University of Hong Kong
Shatin, New Territories, Hong Kong
ph: 852 2609 6862
fx: 852 2603 5723
email: tonyjames@cuhk.edu.hk
.....................................
"nothing great was achieved without enthusiasm."
R. W. Emerson
From pinkert at cmed.bhs.uab.edu Sat Sep 14 09:48:18 1996
From: pinkert at cmed.bhs.uab.edu (Dr. C.A. Pinkert)

Subject: Caesarean sections
Message-ID: <19960914135340470.AAA136@cap1>

> Has anyone any experience with injecting mice with hormones( Day,
> Dose) few Days prior doing caesarean section to avoid any early
delivery.
> I know that the gestation period for the transgenic line I am
> interested in is 19 Days. I planed to do sections on Day 18 p. c..
If the line uniformly delivers on d19 and the pups are robust, then
C-sections on d18 should likely work out well without the additional
hormone therapy.



From mlm at titus.u-strasbg.fr Tue Sep 17 16:19:26 1996
From: mlm at titus.u-strasbg.fr (Marianne LEMEUR)

Subject: zygote yields
Message-ID: <v01510103ae646c5d8502@[130.79.78.57]>

Thanks to those who gave the solution for NOD mice zygote yield!
More generally it seems that C57Bl6 is the only inbred strain which can
be
superovulated in the same way as an F1 hybrid.
Other inbred strains like FVB/N or outbred strains like CD1 do not give
better yields than can be achieved with a natural mating.Is that
correct?
Marianne Le Meur




From jparkert at magnus.acs.ohio-state.edu Tue Sep 17 15:45:24 1996
From: jparkert at magnus.acs.ohio-state.edu (Janice Parker-Thornburg)

Subject: zygote yields
Message-ID: <199609171445.KAA19688@postbox.acs.ohio-state.edu>

>Thanks to those who gave the solution for NOD mice zygote yield!
>More generally it seems that C57Bl6 is the only inbred strain which
can be
>superovulated in the same way as an F1 hybrid.
>Other inbred strains like FVB/N or outbred strains like CD1 do not
give
>better yields than can be achieved with a natural mating.Is that
correct?
>Marianne Le Meur

Marianne:
This is not correct. I have superovulated FVB/N for microinjection, as
well as for general harvesting of mouse embryos. If done in the
correct
time frame, one can get 8-10 more eggs (embryos) per female than with
natural matings. I also know that people using CD1's for embryo
harvesting
will superovulate them, presumably because they get a better yield. If
you
find that your yield does not improve with superovulation, it is likely
that your mouse-age or timespan for the hormone injections needs to be
optimized.

Jan Parker-Thornburg

Jan Parker-Thornburg, Manager
Transgenic Animal Facility
The Ohio State University
jparkert@magnus.acs.ohio-state.edu




From landel at medsci.udel.edu Tue Sep 17 16:37:11 1996
From: landel at medsci.udel.edu (Carlisle Landel)

Subject: zygote yields
Message-ID: <199609171537.LAA03938@helios.medsci.udel.edu>

>Thanks to those who gave the solution for NOD mice zygote yield!
>More generally it seems that C57Bl6 is the only inbred strain which
can be
>superovulated in the same way as an F1 hybrid.
>Other inbred strains like FVB/N or outbred strains like CD1 do not
give
>better yields than can be achieved with a natural mating.Is that
correct?
>Marianne Le Meur

Marianne,

No, this isn't correct! I superovulate FVB/N routinely, and I'm pretty
sure that colleagues of mine in the past have done it to CD1's, too,
though
I vaguely recall there was some trick with dosage or timing or
something
(but I may be wrong--this was many years ago).

Check your hormones for proper concentration, and check that the light
controls in your facility are properly set, and make sure that you
are injecting at or very near the middle of your light cycle.

Regards,

Carlisle Landel
Dept. of Clinical Science
A.I. duPont Institute
PO Box 269
Wilmington DE 19899
(302) 651-6873
landel@helios.medsci.udel.edu
http://mdblmac.medsci.udel.edu/WEB/MDBL/Carlisle.html



From pinkert at cmed.bhs.uab.edu Tue Sep 17 10:23:08 1996
From: pinkert at cmed.bhs.uab.edu (Dr. C.A. Pinkert)

Subject: zygote yields
Message-ID: <19960917142758010.AAB158@cap1>

> More generally it seems that C57Bl6 is the only inbred strain which
can be
> superovulated in the same way as an F1 hybrid.
I'm not sure what is meant here, but other inbred strains can be
utilized, using the same general scheme and sometimes jogging the
schedule a bit.

> Other inbred strains like FVB/N or outbred strains like CD1 do not
give
> better yields than can be achieved with a natural mating.Is that
correct?
It's a matter of degree, but I disagree with the last statement
(unless you mean that hybrids provide better net yields than either
inbreds and outbreds, which is true). In some labs, natural matings
facilitate a specific routine. However, there are significant
economies-of-scale when superovulation is employed (in relation to
zygote yields and needed labor/space/etc.) when using the strains
that you have mentioned.



From browng at medicine.wustl.edu Tue Sep 17 19:03:45 1996
From: browng at medicine.wustl.edu (Gary Brown)

Subject: zygote yields
In-Reply-To: <v01510103ae646c5d8502@[130.79.78.57]>
Message-ID: <Pine.GSO.3.94.960917125610.29137A-100000@medicine>



On Tue, 17 Sep 1996, Marianne LEMEUR wrote:

> Thanks to those who gave the solution for NOD mice zygote yield!
> More generally it seems that C57Bl6 is the only inbred strain which
can be
> superovulated in the same way as an F1 hybrid.
> Other inbred strains like FVB/N or outbred strains like CD1 do not
give
> better yields than can be achieved with a natural mating.Is that
correct?
> Marianne Le Meur
>
>
>
>
Not so. I used FVB/N strain mice for 3 1/2 years and superovulated with
(in my opinion) a high degree of success. Since it's a larger mouse
than
B6C3F1 hybrids, we used 7.5IU hormones rather than 5IU, and a 47 hr gap
between PMSG and HCG. Typical yields were 25-30 good (non fragmented)
eggs per mouse. Can't say I've had experience with other strains other
than C57Bl/6 and B6C3F1 hybrids. Actually, I lie. 129SvJ mice are
dismal
for superovulation - if anyone can give me a way to do these
reproducibly
I'll get you a beer (or three)!

Gary Brown
E-mail: browng@medicine.wustl.edu or 75017.1050@compuserve.com
Web: http://ourworld.compuserve.com/homepages/TheBroons/
         "The Microinjection Workshop"




From carboni at valoritech.com Tue Sep 17 20:25:09 1996
From: carboni at valoritech.com (Nick Carboni)

Subject: Wood's coculture technique
Message-ID: <199609171925.PAA25906@nash.pubnix.net>

Hi everybody,


Short message to know if one of you is currently working or has tried
to
work on wood's coculture technique.

I have spoke with some people, and it appears that it is rather
difficult to
acheive.

Has anyone any experience to share with me ??

Thanks in advance,

Nick
------------------------------------------------------------------
Nick Carboni, VP
Valoritech
1253, McGill College Av., room 620
Montreal, Quebec, Canada
H3B 2Y5
Tel.: (514) 866-8271 / Fax: (514) 866-8272
E-Mail: carboni@valoritech.com
From browng at medicine.wustl.edu Wed Sep 18 13:26:12 1996
From: browng at medicine.wustl.edu (Gary Brown)

Subject: Wood's coculture technique
In-Reply-To: <199609171925.PAA25906@nash.pubnix.net>
Message-ID: <Pine.GSO.3.94.960918070739.1409A-100000@medicine>



On Tue, 17 Sep 1996, Nick Carboni wrote:

> Hi everybody,
>
>
> Short message to know if one of you is currently working or has tried
to
> work on wood's coculture technique.
>
> I have spoke with some people, and it appears that it is rather
difficult to
> acheive.
>
> Has anyone any experience to share with me ??
>
> Thanks in advance,
>
> Nick
>
Hi !

I tried Wood's co-culture technique around 3 years ago, but only had 1
recorded birth in 16 implanted mothers, and this pup was malformed. We
were unable to determine the efficacy of the technique as the ES cell
line
that we used was obtained from another laboratory, and both their
technique and the cell line itself were called into question. However,
here's what I can tell you..

1. We found that the ES cells did not adhere to the de - ZP'd morulae /
octoploid / tetraploid embryos as well as we thought the paper
indicated.

2. Our understanding is that the technique's success is very ES cell
dependent.

3. Our understanding is that the live birth rate is very low, making
this
a system better suited to in utero developmental studies.

Since this time, I have tried to find the time to explore non-injection
methods of chimera production (with little success on the time front!).
I
have only managed to complete a pilot study of Nagy / Rossant's
"sandwich"
method where I was successful in fusing 2 morulae together in culture
overnight in custom made dimples. This effort would have been more
instructive if I had had access to some ES cells to "sandwich", but the
fusions had a >90% success rate. For the reference on this
methodology,
do a Medline search on the authors above or check out the E-newsletter
archive on my homepage for August / September.

Hope this is instructive!     :-)

Gary Brown
E-mail: browng@medicine.wustl.edu or 75017.1050@compuserve.com
Web: http://ourworld.compuserve.com/homepages/TheBroons/
            "The Microinjection Workshop"




From ldeg at midway.uchicago.edu Thu Sep 19 02:28:26 1996
From: ldeg at midway.uchicago.edu (Linda Degenstein)

Subject: ES Cells from Balb C mice
Message-ID: <199609190128.UAA22947@midway.uchicago.edu>

   We are looking for ES Cells (germline proven) that are made from
Balb C
strain mice. Does anyone know if these are available?

Linda Degenstein
Director UCCRC
Transgenic/ES Cell Facility
The University of Chicago
ldeg@midway.uchicago.edu




From khelmuth at facstaff.wisc.edu Tue Sep 24 20:28:52 1996
From: khelmuth at facstaff.wisc.edu (khelmuth@facstaff.wisc.edu)

Subject: noise problems
Message-ID: <v02140b01ae6d9fec8a48@[144.92.94.130]>

Hi everyone:

I am looking for documentation that excess noise can affect the
breeding
and production of embryos from mice. Does anyone know of published
work
that looked at environmental affects on a mouse colony? Any citations
would be appreciated.

Kathy

Kathy Krentz Helmuth
Transgenic Facility
425 Henry Mall
Room 2210 Biotechnology Center
Madison, WI 53706
(608) 265-2801
email:khelmuth@facstaff.wisc.edu




From mlp at informatics.jax.org Tue Sep 24 22:20:46 1996
From: mlp at informatics.jax.org (Moyha Lennon-Pierce)

Subject: Noise/reproduction
Message-ID: <v01540b05ae6dfd6df569@[192.233.41.18]>

The following references might be useful.

I searched the Strain Characteristics Catalog in development at The
Jackson
Lab and did a quick search in NLM. Not much at all in the literature,
but
the following references may be useful.

Nawrot PS et al.
Embryotoxicity of various noise stimuli in the mouse.
Teratology 1980; 22(3):279-89.
CF-1

Zakem HB, Alliston CW.
The effects of noise level and elevated ambient temperatures upon
selected
reproductive traits in female Swiss-Webster mice.
Lab Anim Sci 1974; 24:469-75.
Strain SWR

Moyha Lennon-Pierce
MGD Informatics
mlp@informatics.jax.org

>Hi everyone:
>
>I am looking for documentation that excess noise can affect the
breeding
>and production of embryos from mice. Does anyone know of published
work
>that looked at environmental affects on a mouse colony? Any citations
>would be appreciated.
>
>Kathy
>
>Kathy Krentz Helmuth
>Transgenic Facility
>425 Henry Mall
>Room 2210 Biotechnology Center
>Madison, WI 53706
>(608) 265-2801
>email:khelmuth@facstaff.wisc.edu
From landel at medsci.udel.edu Tue Sep 24 22:13:36 1996
From: landel at medsci.udel.edu (Carlisle Landel)

Subject: Noise/reproduction
Message-ID: <199609242113.RAA08916@helios.medsci.udel.edu>

Kathy,

I saw a poster at a meeting a couple of weeks ago where they were using
one of those ultrasonic mouse repellers to increase the percentage
of embryo loss in CBAxDBA/2 model. If you want more info, I'll see if
I can point you towards the authors.

Regards,

Carlisle Landel
Dept. of Clinical Science
duPont Hospital for Children
PO Box 269
Wilmington DE 19899
(302) 651-6873
landel@helios.medsci.udel.edu
http://mdblmac.medsci.udel.edu/WEB/MDBL/Carlisle.html



From Roger.Leemann at imr.psi.ch Wed Sep 25 15:40:47 1996
From: Roger.Leemann at imr.psi.ch (Roger C. Leemann)

Subject: noise problems
Message-ID: <324941f920c1002@pss200.psi.ch>

Kathy Krentz Helmuth (khelmuth@facstaff.wisc.edu) wrote:

>I am looking for documentation that excess noise can affect the
breeding
>and production of embryos from mice.

Kathy

Sorry, no literature, but I can tell you how we do it. In our animal
housing facility there is always a radio playing at a low volume during
the
daylight cycle (maybe this can not be referred to as "excessive"
noise.) I
don't know whether or how this affects the mice, positive or negative.
With
non-superovulated B6C3F1 mice I can expect 15% to 25% plugs and 7 to 12
two-cell embryos/mouse. How does this compare to other labs?

Roger

:::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::
:::::
Work: Institute for Medical Radiobiology         ++41 (56) 310-3792
[desk]
       CH-5232 Villigen PSI                      ++41   (1) 3856-561
[desk ZH]
       Switzerland                               ++41 (56) 310 3294
[fax]
Home: Nordstrasse 26, CH-8006 Zuerich
       Switzerland                               ++41   (1) 361 0349
[home]



1996 October
From pasceri at sickkids.on.ca Tue Oct 1 17:34:58 1996
From: pasceri at sickkids.on.ca (Peter Pasceri)
Date: Wed Aug 20 11:52:53 2003
Subject: Cryopreservation
Message-ID: <v01510100ae772f68646f@[142.20.24.42]>

Hi all!

I am looking for suppliers of mouse embryo freezing kits. Can you help?

Thanks,

Peter

--
Peter Pasceri
pasceri@sickkids.on.ca
Genetics Research                                         Phone:     (416)
813-4205
The Hospital for Sick Children                            Toronto,
Ontario,Canada




From p.sobieszczuk at ic.ac.uk Thu Oct 3 12:37:29 1996
From: p.sobieszczuk at ic.ac.uk (Peter Sobieszczuk)
Date: Wed Aug 20 11:52:53 2003
Subject: Transgenic mice/Transplantation (fwd)
Message-ID: <v01510104ae7954f36393@[155.198.45.54]>

Sorry, if it is a duplication for some of you.
Peter (your listowner)


>Hello everyone,
>
>Any advise/suggestions on the following questions would be greatly
>appreciated.
>
>Thank you
>
>Anna V. Anagnostopoulos
>TBASE
>Division of Biomedical Information Sciences
>The Johns Hopkins University School of Medicine
>anna@gdb.org
>
>---------------
>
>I am working with mice, doing Bone Marrow and Peripheral Blood
>Tranplantation experiments, at the Department of Immunogenetics of the
New
>York Blood Center. We are interested in developing a syngeneic mouse
>transplantation model. We would like to use as donors, animals
carrying a
>transgene, that its product is not lethal or significantly affecting
the
>recipients (especially hematologically), and also is easily detectable
>post-transplant. For example, mice transgenic for a human gene,
encoding
>for a human antibody, would be a good model. The idea would be to show
>that the donor cells (positive for the transgene) engraft, and the
>transgene is functional (we can detect the product) after transplant.
The
>mouse strains I am working with at the present time are C57BL/6, A/J
and
>B6A (C57BL/6 X A/J) (all obtained from Jackson Labs). It would help
>tremendously if the transgenic animals are of the same background. Do
you
>have any suggestions?
>




From mangia at uniroma1.it Fri Oct 4 13:46:36 1996
From: mangia at uniroma1.it (Franco Mangia)
Date: Wed Aug 20 11:52:53 2003
Subject: Transgenic mice/Transplantation (fwd)
Message-ID: <E0v98iC-0006aA-00@juliet.ic.ac.uk>


>
>We are interested in developing a syngeneic mouse
>>transplantation model. We would like to use as donors, animals
carrying a
>>transgene, that its product is not lethal or significantly affecting
the
>>recipients (especially hematologically), and also is easily
detectable
>>post-transplant. For example, mice transgenic for a human gene,
encoding
>>for a human antibody, would be a good model. The idea would be to
show
>>that the donor cells (positive for the transgene) engraft, and the
>>transgene is functional (we can detect the product) after transplant.
The
>>mouse strains I am working with at the present time are C57BL/6, A/J
and
>>B6A (C57BL/6 X A/J) (all obtained from Jackson Labs). It would help
>>tremendously if the transgenic animals are of the same background. Do
you
>>have any suggestions?
>>
>Anna V. Anagnostopoulos
>TBASE
>Division of Biomedical Information Sciences
>The Johns Hopkins University School of Medicine
>anna@gdb.org



I think that the ROSA26 transgenic mice, developed by Phil Soriano, are
just what you need for labeling engrafts at a single cell level. These
mice carry a lacZ transgene directed by a ubiquitous promoter active in
all
tissues, without apparently affecting cell viability, since these mice
are
fine. These mice can be obtained from the Jackson Labs either as
B6,129 F1
(the original Soriano's hybrid strain) or with a pure C57BL/6J
background.
Complete names of these mice are: "B6,129-TgR(ROSA26)26 Sor", and
"C57Bl/6J-TgR(ROSA26)26 Sor".   I am also very interested in obtaining
an
ES cell line(s) from these mice. Is anybody aware whether these ES
lines
are being developed in some lab? Also, does anybody know whether
ROSA26
mice with nuclearly targeted beta-gal have been produced?
Franco



Franco Mangia
Laboratory of General Biology
Department of Psychology
University La Sapienza of Rome
c/o Institute of Histology and General Embryology
Via Borelli, 50
00161 Rome, Italy
tel: 39-6-4976-8103   FAX: 39-6-4976-8099
E-mail: mangia@uniroma1.it




From p.sobieszczuk at ic.ac.uk Sat Oct 5 19:19:10 1996
From: p.sobieszczuk at ic.ac.uk (Peter Sobieszczuk)
Date: Wed Aug 20 11:52:53 2003
Subject: "TgList Backstage" (admin. news *2) ARCHIVES & TBASE
Message-ID: <v01510102ae7c443e5426@[155.198.45.54]>

Dear Transgenic Subscribers,

Greetings to newcomers,

ARCHIVES
Charmaine Foltz asked me some time ago:
>I was wondering if the information on the list will be retrievable
>(i.e., will there be a data base that we can search archived files).

very much so, send the message:
                index transgenic-list

to listserver at
                   majordomo@ic.ac.uk

and you shall receive an index of monthly files, something like:

>>>> index transgenic-list
total 241
-rw-rw---- 1 daemon       10364    Jul 29 16:38 transgenic-
list.archive.9607
-rw-rw---- 1 majordom    146666    Aug 29 21:36 transgenic-
list.archive.9608
-rw-rw---- 1 majordom     71873    Sep 25 15:31 transgenic-
list.archive.9609
-rw-rw---- 1 majordom      6715    Oct   4 12:46 transgenic-
list.archive.9610
>>>>

where July entries file name is "transgenic-list.archive.9607"

If you then send:
        get transgenic-list transgenic-list.archive.9607

you should receive all postings for that month (in this case, July) as
individual e-mail messages.

If you have access to world wide web, (WWW) you can explore archives
with
the luxury of selecting individual entries etc., using your favourite
browser, Netscape, Mosaic or Explorer.

The URL address is:
        http://www.lists.ic.ac.uk/hypermail/transgenic-list

I'm afraid however that even with the browser, files are not searchable
(at
this stage anyway), although subject, (which should be the topic of any
message) is clearly visible.


TBASE
Anna Anagnostopoulos who is co-ordinating TBASE data base site sent
this
introduction (with minor editorial changes P.S.):
-----------------------------------------------------------------------
-
BEGINNING OF TEXT

TBASE, the Transgenic/Targeted Mutation Database, is an attempt to
organize information on transgenic animals and targeted mutations
generated, and analyzed worldwide. TBASE is available on the WWW

http://www.gdb.org/Dan/tbase/tbase.html

and can be searched either
quite broadly across the entire database (the 'blunt object' approach)
or
quite specifically with the use of 28 distinct searchable fields (the
'scalpel and tweezers' approach). TBASE contains a wealth of
information
about each animal in the database. A selection of the fields present in
the databases are: line name, genus, DNA construct description, host
background, line source, method used , transgene/targeted allele
expression, lethality, phenotype, handling, contact information, full
citation and abstracts of the papers related to the creation of a line.

Owing to the wide use of homologous recombination in ES cells,
TBASE initially focused on maintaining a comprehensive coverage of
all targeted mutations reported to date. Moreover, it focused on the
mouse
as the predominant mammalian model. Specific emphasis is still placed
on
cataloging novel knockout mice, which have defined genetic
modifications,
with an extensive detailed description of their resulting mutant
phenotypes. In the future, the focus of TBASE will expand to cover
all animals resulting from transgenesis or targeted mutation,
regardless
of species. Additionally, TBASE plans to incorporate 'transgenic
knockouts', that is, knockout animals that have subsequently served as
recipients for transgenes.

TBASE should be viewed as a dynamic database with a potentially
unlimited
number of entries, expected to grow considerably as more experimental
methods and lines are generated. Data acquisition is primarily effected
by
active literature scanning and manual data entry, as well as processing
of
direct submission forms.

Literature scanning refers to direct data extraction from the
scientific
literature through regular examination of over 20 journals. These
journals have been statistically identified as periodically stable
sources
of information on transgenesis and gene targeting. Active literature
scanning has, so far, been the primary mode of data accumulation, and
has
ensured that the database faithfully reflects the general direction of
related research. It is expected to remain a significant component of
the
data acquisition tactics, even as other methods of data acquisition
become
available (see below). In an effort to avoid missing important data,
keyword-related references from additional journals are also screened
by
the use of Medline and Current Contents.

In the future, data that are not manually entered by TBASE staff
through the literature scanning process, are expected to arrive either
as
direct paper or as electronic submissions. Although over 98% of the
data
have been entered manually so far, direct paper submissions are
expected
to increase as TBASE becomes fully adopted by the scientific
community. In addition, as electronic submission tools become
available,
more research groups will be prepared to commit resources to the
production of data deposited into TBASE. Electronic submission
tools and their error-checking capabilities will ensure efficient and
timely deposition of the data.

END OF TEXT
-----------------------------------------------------------------------
-----


STATISTIC:
There are approx. 240 members from at least 18 countries registered
now,
and I'm starting to think aren't we growing too big, read to much to
read.

Regards,

Yours sincerely,

Listowner




_______________________________________________________________________
____

Dr Peter Sobieszczuk
Imperial College School of Medicine at St. Mary's        tel: 0171 594
3784
Biochemistry and Molecular Genetics                      tel: 0171 594
3799
Norfolk Place                                            fax: 0171 706
3272
London W2 1PG, U.K.                          e-mail:
p.sobieszczuk@ic.ac.uk
_______________________________________________________________________
____




From khelmuth at facstaff.wisc.edu Wed Oct 9 10:28:47 1996
From: khelmuth at facstaff.wisc.edu (khelmuth@facstaff.wisc.edu)
Date: Wed Aug 20 11:52:53 2003
Subject: fire codes within animal colony
Message-ID: <v02140b00ae7ef3eb32c0@[144.92.94.130]>

Hello everyone-

I face a delemma within our facility and am hoping someone can help me
out.
We recently moved our rat and mouse colonies into a new building.
Everything seems to be going well except for one thing. We experienced
the
first fire drill and realized that there are 3 fire alarms within our
5ft.
wide clean hallway and 3 fire alarms within our 5ft. wide dirty
hallway.
They are extremely loud and potentially problematic for our animals.
Our
building manager agrees that this seems extremely overdone and wishes
to
get some of the alarms diconnected. However, the state fire code
regulator
feels that with the background noises of the cage washer, movement of
carts
and racks, etc. that these extra fire alarms are necessary. He will
only
follow suit if I can come up with evidence that not only does it affect
the
well-being of the animals but that other facilities have a small number
of
alarms. I have obtained documentation of loud noises and it's effect
on
breeding, embryo production, etc. Can someone help me out with
regulations
regarding fire alarms within your animal facility?

Any suggestion on how to deal with this would be beneficial.
Thanks

Kathy Krentz Helmuth

Kathy Krentz Helmuth
Transgenic Facility
425 Henry Mall
Room 2210 Biotechnology Center
Madison, WI 53706
(608) 265-2801
email:khelmuth@facstaff.wisc.edu




From landel at medsci.udel.edu Wed Oct 9 11:44:03 1996
From: landel at medsci.udel.edu (Carlisle Landel)
Date: Wed Aug 20 11:52:53 2003
Subject: fire codes within animal colony
Message-ID: <199610091444.KAA19359@helios.medsci.udel.edu>

I'm curious--how often do these alarms go off? Maybe instead you could
get them to absolve you from frequent fire drills.

Carlisle Landel

>From owner-transgenic-list@ic.ac.uk Wed Oct 9 10:35:44 1996
>Mime-Version: 1.0
>Content-Type: text/plain; charset="us-ascii"
>To: transgenic-list@ic.ac.uk
>From: khelmuth@facstaff.wisc.edu
>Subject: fire codes within animal colony
>Sender: owner-transgenic-list@ic.ac.uk
>Precedence: bulk
>Reply-To: transgenic-list@ic.ac.uk
>
>Hello everyone-
>
>I face a delemma within our facility and am hoping someone can help me
out.
>We recently moved our rat and mouse colonies into a new building.
>Everything seems to be going well except for one thing. We
experienced the
>first fire drill and realized that there are 3 fire alarms within our
5ft.
>wide clean hallway and 3 fire alarms within our 5ft. wide dirty
hallway.
>They are extremely loud and potentially problematic for our animals.
Our
>building manager agrees that this seems extremely overdone and wishes
to
>get some of the alarms diconnected. However, the state fire code
regulator
>feels that with the background noises of the cage washer, movement of
carts
>and racks, etc. that these extra fire alarms are necessary. He will
only
>follow suit if I can come up with evidence that not only does it
affect the
>well-being of the animals but that other facilities have a small
number of
>alarms. I have obtained documentation of loud noises and it's effect
on
>breeding, embryo production, etc. Can someone help me out with
regulations
>regarding fire alarms within your animal facility?
>
>Any suggestion on how to deal with this would be beneficial.
>Thanks
>
>Kathy Krentz Helmuth
>
>Kathy Krentz Helmuth
>Transgenic Facility
>425 Henry Mall
>Room 2210 Biotechnology Center
>Madison, WI 53706
>(608) 265-2801
>email:khelmuth@facstaff.wisc.edu
>
>
>
>
>



From duffyh at war.wyeth.com Wed Oct 9 15:45:06 1996
From: duffyh at war.wyeth.com (Heidi Duffy)
Date: Wed Aug 20 11:52:53 2003
Subject: fire codes within animal colony -Reply
Message-ID: <s25bb9fa.076@war.wyeth.com>

I used to work in animal facility that used flashing lights instead of
 audible alarms within the animal facility. The audible alarms were
only in the office areas of the facility. (Our phones within the
facility were set up the same way, including the cagewash areas).
Designated techs were assigned to check all animal rooms for
personnel evacuation before leaving the buildings. It worked out very
well.

Heidi Duffy
email:duffyh@war.wyeth.com




From david_gask at Merck.Com Thu Oct 10 13:47:00 1996
From: david_gask at Merck.Com (David Gask)
Date: Wed Aug 20 11:52:53 2003
Subject: Fire Alarm problems-Kathy Krentz Helmuth
Message-ID: <199610101150.HAA21899@igw2>


Kathy,
              Why don't you investigate the use of ' SILENTONE ALARMS'
as
an alternative. These are used widely in the UK and were developed to
prevent problems for the animals caused by more conventional sounders.
They
produce a loud noise suitable for use as an alarm in the event of fire,
but
at the same time the frequency is below the threshold frequency, or
outside
the most sensitive range for most commonly used laboratory animals. If
you
require the address of a supplier I can send it to you. All our animal
facility fire alarms are of this type.

Hope this helps

Dave Gask
Operations Coordinator
Merck Sharp & Dohme
Neuroscience Research Centre
Terlings Park
Eastwick Road
Harlow, Essex
CM20 2QR
England
Tel 01279 440220
e-mail David_Gask@Merck.com


Hello everyone-

I face a delemma within our facility and am hoping someone can help me
out.
We recently moved our rat and mouse colonies into a new building.
Everything seems to be going well except for one thing. We experienced
the
first fire drill and realized that there are 3 fire alarms within our
5ft.
wide clean hallway and 3 fire alarms within our 5ft. wide dirty
hallway.
They are extremely loud and potentially problematic for our animals.
Our
building manager agrees that this seems extremely overdone and wishes
to
get some of the alarms diconnected. However, the state fire code
regulator
feels that with the background noises of the cage washer, movement of
carts
and racks, etc. that these extra fire alarms are necessary. He will
only
follow suit if I can come up with evidence that not only does it affect
the
well-being of the animals but that other facilities have a small number
of
alarms. I have obtained documentation of loud noises and it's effect
on
breeding, embryo production, etc. Can someone help me out with
regulations
regarding fire alarms within your animal facility?

Any suggestion on how to deal with this would be beneficial.
Thanks
Kathy Krentz Helmuth



From Roger.Leemann at imr.psi.ch Thu Oct 10 16:21:53 1996
From: Roger.Leemann at imr.psi.ch (Roger C. Leemann)
Date: Wed Aug 20 11:52:53 2003
Subject: Rodent-research list
Message-ID: <v01540b00ae82a600b461@[129.129.90.92]>

Hi

Since I'm working with mice but do not use transgenic animals, I
subscribed
to the rodent-research list about a month ago. Majordomo told me, that
it
had forwarded my request to the list owner for approval but I haven't
got
any response since.
Maybe it's not a list for every Jack and Joe (or was it Jack and
Jill?), or
maybe the list owner is too busy, or on vacation, whatever. I don't
mind if
they don't want me, but ones likes to know...
Anybody in this list who has a clue why the rodent-research list seems
dead?

Roger

:::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::
:::::
Work: Institute for Medical Radiobiology        ++41 (56) 310-3792
[desk]
       CH-5232 Villigen PSI                     ++41 (1) 3856-561
[desk ZH]
       Switzerland                              ++41 (56) 310 3294
[fax]
Home: Nordstrasse 26, CH-8006 Zuerich
       Switzerland                              ++41 (1) 361 0349
[home]




From p.sobieszczuk at ic.ac.uk Thu Oct 10 17:55:30 1996
From: p.sobieszczuk at ic.ac.uk (Peter Sobieszczuk)
Date: Wed Aug 20 11:52:53 2003
Subject: Rodent-research list & "TgL Backstage"(admin.news*3)
Message-ID: <v01510100ae82cd2a148c@[155.198.45.54]>

Dear Roger,

>Since I'm working with mice but do not use transgenic animals, I
subscribed
>to the rodent-research list about a month ago. Majordomo told me, that
it
I presume listserver "majordomo" at Caltech, and not "ic.ac.uk" at
Imperial College?

>Maybe it's not a list for every Jack and Joe (or was it Jack and
Jill?), or
Do not take it personnally, it's just a computer.

>maybe the list owner is too busy, or on vacation, whatever. I don't
mind if
>they don't want me, but ones likes to know...

Sorry, I still do not know what exactly happened, Eric Mercer had some
technical problems supporting "rodent-research" list, 3 or 4 months
ago. A few people has tried to contact him, at the end, this list was
created (see the introduction to the list available also by sending
"info transgenic-list" message to "majordomo@ic.ac.uk" address).

You have been with us (transgenic-list) for the month now and probably
noticed that the list is un-moderated and has no restriction on the
content. Transgenic - is just a name on a server, and the list a quick
way to exchange information.

I would not like to start a discussion on definition of transgenesis,
but would imagine that for the active researchers in the field it would
cover quite a bit of biology, from molecular to developmental, with the
animal models not exclussively restricted to rodents.

So it is not the name but participants who will take the discussion in
any particular direction, including yourself.

Who would you like to see on the list Roger, or should I ask everybody:

Who else do you think we should invite to the list?

Best regards,

Sincerely,

Peter - your listowner




From s.tan at anatomy.unimelb.edu.au Tue Oct 15 16:09:53 1996
From: s.tan at anatomy.unimelb.edu.au (S.Tan)
Date: Wed Aug 20 11:52:53 2003
Subject: Fostering baby mice
Message-ID: <v03007812ae88bef7a8eb@[128.250.225.71]>

I am grafting pieces of neural tissue into the cortex of newborn mice
but
find that the mother keeps eating the babies. The wound is closed with
a
single silk suture and this appears to attract the attention of mum.
The
mother's strain is C57/BL6 x DBA. Are there less aggressive strains of
mums out there? Thanks.




From a.annala at ucl.ac.uk Tue Oct 15 06:51:24 1996
From: a.annala at ucl.ac.uk (Dr. Alexander J. Annala)
Date: Wed Aug 20 11:52:53 2003
Subject: Fostering baby mice
Message-ID: <v01540b00ae88e518e7ca@[128.40.81.204]>

>I am grafting pieces of neural tissue into the cortex of newborn mice
but
>find that the mother keeps eating the babies. The wound is closed
with a
>single silk suture and this appears to attract the attention of mum.
The
>mother's strain is C57/BL6 x DBA. Are there less aggressive strains
of
>mums out there? Thanks.

Most F1's make very good mothers. Your C57BL/6 x DBA are F1 mice.
We use C57BL/6 x CBA and C57BL/10 x CBA with good results.

Do you take precaution to avoid leaving scent on operated baby mice?
Some people roll babies in mom's urine before returning them to the
nest. Others treat mom's nose with alcohol to keep her from smelling
any differences between babies for awhile. This might be a scent issue
rather than an exposed suture problem.

Internalize suture might be difficult in baby mice. Maybe you could
make your incision somewhere away from your skull opening, slide
the skin opening over the area you want to open bone, do your work,
slide the skin back, and maybe use one of the tissue adhesives which
are pretty available around medical schools and operating theatres
to close the wound. This would avoid leaving exposed suture for the
mom to chew on.




From fobo at ltk.unizh.ch Tue Oct 15 08:22:40 1996
From: fobo at ltk.unizh.ch (Frank Bootz)
Date: Wed Aug 20 11:52:53 2003
Subject: Fostering baby mice
Message-ID: <1.5.4.32.19961015062240.0068aa74@rzu-mailhost.unizh.ch>

Dear colleaque,
I have done thymectomy in newborn mouse. The wound was also closed with
a
single U-suture in the neck. The pups awake within 5 minutes after
methofane
(methoxyflurane) anesthesia and brought back to the mother immediately.
We
don't anesthetize the mother, contrary the most papers describing this
procedure. In this case the mother's strain is C57/Bl/6 x PO. If we can
choose the strain, we normally use Zur:ICR original Charles River swiss
mice
CD-1. They are mice with excellent mother attributes.

Best regards,

Frank
Frank Bootz
Institute of Laboratory Animal Science
University of Zurich


Winterthurerstrasse 190                     phone: +41 1 257 54 55
8057 Zurich                                 fax:   +41 1 257 57 03
Switzerland                                 e-mail fobo@ltk.unizh.ch




From GWOLFF at NCTR.FDA.GOV Tue Oct 15 08:44:51 1996
From: GWOLFF at NCTR.FDA.GOV (George L. Wolff)
Date: Wed Aug 20 11:52:53 2003
Subject: Fostering baby mice
Message-ID: <695B452EEE@EFS.NCTR.FDA.GOV>

Zur:ICR cannot be descended from CD-1 mice - or the strain
designation is incorrect. Let me know if you'd like more details.
George L. Wolff



Tel: (501) 543-7522
FAX: (501) 543-7635/7662
NCTR       HFT-140
3900 NCTR Road
Jefferson, AR 72079-9502




From gotto at leland.Stanford.EDU Wed Oct 16 13:03:00 1996
From: gotto at leland.Stanford.EDU (Glen Otto)
Date: Wed Aug 20 11:52:53 2003
Subject: Fostering baby mice
In-Reply-To: <1.5.4.32.19961015062240.0068aa74@rzu-mailhost.unizh.ch>
Message-ID: <v03007800ae8adbf9a0a6@[171.65.18.36]>

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From a.annala at ucl.ac.uk Wed Oct 16 22:50:59 1996
From: a.annala at ucl.ac.uk (Dr. Alexander J. Annala)
Date: Wed Aug 20 11:52:53 2003
Subject: Fostering baby mice -- Failure to thrive?
Message-ID: <E0vDcxZ-0002Gs-00@juliet.ic.ac.uk>

What can be done to assist baby mice in distress after birth? Some
chimeras with neuroreceptor channel mutations experience difficulty
feeding themselves -- and sometimes even suckling milk from mother.

We have in the past injected approx 20% weight of animal with Ringers
Lactate (Hartman's Solution). However there must be more aggressive
therapy which would facilitate survival of distressed baby mice.

We have also used incubator -- with reservations about the possibility
of dehydration from elevated environmental temperature.

Thanks,

Alexander J. Annala, Ph.D.
Senior Research Fellow
Laboratory for Molecular Pharmacology
University College London




From carton at murray.fordham.edu Wed Oct 16 19:44:40 1996
From: carton at murray.fordham.edu (Jill Carton)
Date: Wed Aug 20 11:52:53 2003
Subject: knockout mouse construct problems
Message-ID: <v02130500ae8b106fe16f@[169.132.99.88]>

        We are working on generating several knockout mice and have
been
successful in producing transgenes for some of our genes, but we have a
fairly consistent problem with the final ligation step of assembling
the
construct. When we PCR our ligation reaction to see if the DNA pieces
have
ligated we do get the appropriate product however analysis of the
clones
by miniprep show no positive clones. So we are suspecting that the
problem
is in the uptake or growth of the construct in the bacterial cells.
Although, each fragment of the transgene can be propagated in the
bacteria.
        At this stage we are working with a very large piece of DNA
(~12KB)- and perhaps this will lower the ligation frequency-but we have
generated larger plasmids with little problem.
        We are using JM109 or DH5a as our competent cells (chemically
competent).   Any suggestions at all would be greatly appreciated!

Jill Carton
Dept. Biological Sciences
Fordham University
Larkin Hall, rm 160
Bronx, NY 10458
718-817-3652
718-817-3645 (fax)
carton@murray.fordham.edu




From p.sobieszczuk at ic.ac.uk Thu Oct 17 16:23:53 1996
From: p.sobieszczuk at ic.ac.uk (Peter Sobieszczuk)
Date: Wed Aug 20 11:52:53 2003
Subject: Nod/Scid mice
Message-ID: <v01510103ae8bffb476b7@[155.198.45.54]>

Forwarded from [Anna Anagnostopoulos <anna@screams.gdb.org>]


>Hello everyone,
>
>Dr. Sally J. Cutler (Department of Medical Microbiology, Charing Cross
>Hospital, London. Telephone:+ 44181 846 7570) is looking for sources
of
>Nod/Scid mice, preferably in the UK. Any suggestions on where to
look?
>
>Thank so much
>
>~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
>Anna V. Anagnostopoulos, Ph.D.
>Division of Biomedical Information Sciences
>The Johns Hopkins University School of Medicine
>2024 E. Monument Street
>Baltimore, MD 21205-2236
>tel: (410) 614-3226
>fax: (410) 614-0434
>e-mail: anna@gdb.org




From james at icr.ac.uk Thu Oct 17 18:00:12 1996
From: james at icr.ac.uk (james wallace)
Date: Wed Aug 20 11:52:53 2003
Subject: Nod/Scid mice
Message-ID: <SIMEON.9610171712.A@sbp112.icr.ac.uk>



>
> >Dr. Sally J. Cutler (Department of Medical Microbiology, Charing
Cross
> >Hospital, London. Telephone:+ 44181 846 7570) is looking for
sources of
> >Nod/Scid mice, preferably in the UK. Any suggestions on where to
look?
> >
> >Thank so much
> >
> >~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
> >Anna V. Anagnostopoulos, Ph.D.
> >Division of Biomedical Information Sciences
> >The Johns Hopkins University School of Medicine
> >2024 E. Monument Street
> >Baltimore, MD 21205-2236
> >tel: (410) 614-3226
> >fax: (410) 614-0434
> >e-mail: anna@gdb.org
>
> Hi,
        Tell Sally to give me a call as we have a colony
of Nod\Scids here at ICR in the UK.

Best Wishes


Jim Wallace
Institute of Cancer Research
The McElwain Laboratories
15 Cotswold Rd
Belmont Sutton
Surrey SM2 5NG
England
tel:    0181 643 8901 Ext 4638
Fax:    0181 770 1395
e_mail james@icr.ac.uk
>
>




From anna at screams.gdb.org Thu Oct 17 14:18:07 1996
From: anna at screams.gdb.org (Anna Anagnostopoulos)
Date: Wed Aug 20 11:52:53 2003
Subject: Available enhancer traps
Message-ID: <Pine.SOL.3.91.961017131414.2600G-100000@screams.gdb.org>


Hello again,

If any of you is interested in Joe Miano's enhancer traps or knows of a
bulletin where he can have the pictures posted please let me know.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Anna V. Anagnostopoulos, Ph.D.
Division of Biomedical Information Sciences
The Johns Hopkins University School of Medicine
2024 E. Monument Street
Baltimore, MD 21205-2236
tel: (410) 614-3226
fax: (410) 614-0434
e-mail: anna@gdb.org


---------- Forwarded message ----------
Date: Wed, 16 Oct 1996 16:36:44 -0500
From: Joe Miano <jmiano@post.its.mcw.edu>
To: anna@gdb.org
Subject: enhancer trap


We do transgenics and I have two questions:

(1) Would you or anyone you know be interested in an enhancer trap
mouse
that has expression in a discrete region of each limb as well as the
face?
I hesitate to kill these mice off because I can't help but think
someone,
somewhere might be interested.

(2) Do you know of a bulletin board for posting pictures of enhancer
trap
(gene trap etc) mice? I think such a board would be quite valuable.
For
example, we have genrated several enhancer trap mice (one that stained
a
region of the brain and the spinal cord is regrettably gone). None of
these
mice are of interest to us since we are looking for a SMC-restricted
pattern
of expression (see SM22 transgenic mouse paper in March '96 issue of
JCB).
There may be others, however, who would gladly accept such mice.


Sincerely,

Joseph M. Miano, Ph.D.




From kelly at citi2.fr Thu Oct 17 21:42:58 1996
From: kelly at citi2.fr (U344)
Date: Wed Aug 20 11:52:53 2003
Subject: knockout mouse construct problems
Message-ID: <v02120d03ae8cb9b6d37f@[194.254.89.19]>
>        At this stage we are working with a very large piece of DNA
>(~12KB)- and perhaps this will lower the ligation frequency-but we
have
>generated larger plasmids with little problem.
>        We are using JM109 or DH5a as our competent cells (chemically
>competent). Any suggestions at all would be greatly appreciated!

        Whenever I'm forced to deal with DNA of such a size, I use the
Hanahan transformation proceedure to produce competent cells... the
original reference is J. Mol. Biol. 1983, (166) pg 557... it may also
be
to your advantage to lift and screen colonies before picking them.

                                good luck...

INSERM Unite 344 Endocrinologie Moleculaire
156 rue de Vaugirard, 75730 Paris cedex 15 France

'Les raisonables ont dure, les passiones ont vecu'




From ldeg at midway.uchicago.edu Fri Oct 18 18:19:37 1996
From: ldeg at midway.uchicago.edu (Linda Degenstein)
Date: Wed Aug 20 11:52:53 2003
Subject: Transgenic Facility Yellow Pages
Message-ID: <199610182219.RAA15525@midway.uchicago.edu>

    I am often contacted by people outside my university inquiring as to
whether or not we can produce KO or transgenic mice for them. These are
often from investigators out of the state or even out of the country. I
try
to match them up with an institution in their location if I know of
one.

   I would like to make a list of all the known facilities, with
pertinent
info, such as institution, location, services offered, contact person,
telephone and e-mail listings, and whether or not you can perform these
services for someone outside your institution, but in your area.

   If you are willing to e-mail this info, I will summarize in about a
week
for both lists.

   Thank you for your help.

Linda Degenstein
Director UCCRC
Transgenic/ES Cell Facility
The University of Chicago
ldeg@midway.uchicago.edu




From pinkert at cmed.bhs.uab.edu Fri Oct 18 18:31:33 1996
From: pinkert at cmed.bhs.uab.edu (Dr. C.A. Pinkert)
Date: Wed Aug 20 11:52:53 2003
Subject: Transgenic Facility Yellow Pages
Message-ID: <19961018225449159.AAA83@compaq.uab.edu>

>   Date:        Fri, 18 Oct 1996 17:19:37 -0500 (CDT)
>   To:          transgenic-list@ic.ac.uk, compmed@WUVMD.WUSTL.edu
>   From:        ldeg@midway.uchicago.edu (Linda Degenstein)
>   Subject:     Transgenic Facility Yellow Pages
>   Reply-to:    transgenic-list@ic.ac.uk

>     I am often contacted by people outside my university inquiring as
to
> whether or not we can produce KO or transgenic mice for them. These
are
> often from investigators out of the state or even out of the country.
I try
> to match them up with an institution in their location if I know of
one.
>
>     I would like to make a list of all the known facilities, with
pertinent
> info, such as institution, location, services offered, contact
person,
> telephone and e-mail listings
I think you have info for the NICHD facility at UAB. I'm not sure if
such a listing of all facilities will be problematic with the patent
positions that DNX
(as well as GenPharm, Lexicon, etc. for ES cell work) are staking
out. But so long as the caution is there in advance, it may all be OK.

Carl



From a.annala at ucl.ac.uk Sun Oct 20 14:32:43 1996
From: a.annala at ucl.ac.uk (Dr. Alexander J. Annala)
Date: Wed Aug 20 11:52:53 2003
Subject: Transgenic Facility Yellow Pages
Message-ID: <v01540b00ae8fe7874a90@[128.40.81.204]>

Dr. Pinkert,

I am curious about the patent claims you mention. We do some
pronuclear
microinjection of DNA, ES cell injection into blastocysts, and
aggregation
of ES cells with morula to produce transgenic mice. Are individuals
and/or
organizations claiming patent rights which would preclude carrying out
these basic transgenic procedures in academic laboratories? Are
license
fees to obtain the right to perform these procedures anywhere within
the
reach of small academic laboratories?

Are the basic transgenic patent claims analogous to the original
Stanford
recombinant DNA claims where a $10,000 fee was required for any lab
to perform basic molecular biology techniques? It would seem few, if
any, academic laboratories pay this fee today.

It might be interesting for people to have an up to date listing
available on
the network containing a summary of transgenic relevant patent claims
and licensing arrangements/fees.

Thanks,

Alexander J. Annala, Ph.D.
Wellcome Senior Research Fellow
Laboratory for Molecular Pharmacology
University College London
Gower Street
London WC1E 6BT

Tel: +44(171)380-7857
Fax: +44(171)380-7245
Email: a.annala@ucl.ac.uk

>> Date:           Fri, 18 Oct 1996 17:19:37 -0500 (CDT)
>> To:             transgenic-list@ic.ac.uk, compmed@WUVMD.WUSTL.edu
>> From:           ldeg@midway.uchicago.edu (Linda Degenstein)
>> Subject:        Transgenic Facility Yellow Pages
>> Reply-to:       transgenic-list@ic.ac.uk
>
>>     I am often contacted by people outside my university inquiring as
to
>> whether or not we can produce KO or transgenic mice for them. These
are
>> often from investigators out of the state or even out of the
country. I try
>> to match them up with an institution in their location if I know of
one.
>>
>>     I would like to make a list of all the known facilities, with
pertinent
>> info, such as institution, location, services offered, contact
person,
>> telephone and e-mail listings
>I think you have info for the NICHD facility at UAB. I'm not sure if
>such a listing of all facilities will be problematic with the patent
>positions that DNX
>(as well as GenPharm, Lexicon, etc. for ES cell work) are staking
>out. But so long as the caution is there in advance, it may all be
OK.
>
>Carl




From pinkert at cmed.bhs.uab.edu Sun Oct 20 10:02:00 1996
From: pinkert at cmed.bhs.uab.edu (Dr. C.A. Pinkert)
Date: Wed Aug 20 11:52:53 2003
Subject: Transgenic Facility Yellow Pages
Message-ID: <19961020142523272.AAA152@compaq.uab.edu>

> Dr. Pinkert,
>
> I am curious about the patent claims you mention. We do some
pronuclear
> microinjection of DNA, ES cell injection into blastocysts, and
aggregation
> of ES cells with morula to produce transgenic mice. Are individuals
and/or
> organizations claiming patent rights which would preclude carrying
out
> these basic transgenic procedures in academic laboratories?
If you are providing services in house, there should be no problem.
If you provide services outside your home institution, you can be
liable to the folks holding patents on all the microinjection/gene
transfer patents that have been awarded.
  Are license
> fees to obtain the right to perform these procedures anywhere within
the
> reach of small academic laboratories?
When it is allowed by the patent holders. But if the work is
in-house AND for non-commercial basic research - most of the
companies with the rights will leave you alone. It is only when you
offer a service or begin toward commercialization that you will find
yourself in need of appropriate legal counsel.
>
> Are the basic transgenic patent claims analogous to the original
Stanford
> recombinant DNA claims where a $10,000 fee was required for any lab
> to perform basic molecular biology techniques? It would seem few, if
> any, academic laboratories pay this fee today.
Agreed, but as an example, for DNA microinjection outside of your
home institution, DNX would not allow you to provide the service (as
it competed with their program - but in-house basic research was
automatically exempted). Now, if you want to commercialize or
transfer animals, they have a basic fee structure in mind. However,
if you agree to provide microinjection services to say another
institution - even at/below your actual costs, you would likely be
liable for treble damages . . .
>
> It might be interesting for people to have an up to date listing
available on
> the network containing a summary of transgenic relevant patent claims
> and licensing arrangements/fees.
I agree, but some of the ES cell patents as well as more recent
targeting/expression patents are in process and disclosure is
not readily available.
>
Good luck.

Carl A. Pinkert



From stewarv at cesmtp.ccf.org Mon Oct 21 12:27:16 1996
From: stewarv at cesmtp.ccf.org (Valerie Stewart MS)

Subject: Transgenic Facility Yellow Pages -Reply
Message-ID: <s26b5f4a.015@cesmtp.ccf.org>

Hi Linda--

Good idea, though with the patent battles, our reach is limited. Our
policy so far has been to make mice for Clinic investigators and their
collaborators. Here is the information you requested:

Transgenic/IKnockout Core Facility
Cleveland Clinic Research Institute
FF6-50, 9500 Euclid Avenue
Cleveland, OH 44106
Director, Valerie Stewart
Technician, Christine Brant
phone (216)444-8521
fax (216)445-6257
e-mail "stewarv@cesmtp.ccf.org"
Services include pronuclear injection, blastocyst injection,
cryopreservation by vitrification, training in all techniques. We
also do microinjection of proteins, etc. into cell lines via an
Eppendorf automated system.
Services and prices can also be accessed through the Clinic's web
page at "www.ccf.org". Go to Main Menu, then Research Institute,
then Scientific Support Services, then Core Services.

>>> Linda Degenstein <ldeg@midway.uchicago.edu> - 10/18/96 6:19 PM
>>>
    I am often contacted by people outside my university inquiring as
to whether or not we can produce KO or transgenic mice for them.
These are often from investigators out of the state or even out of
the country. I try to match them up with an institution in their
location if I know of one.

   I would like to make a list of all the known facilities, with
pertinent info, such as institution, location, services offered,
contact person, telephone and e-mail listings, and whether or not you
can perform these services for someone outside your institution, but
in your area.

   If you are willing to e-mail this info, I will summarize in about
a week for both lists.

   Thank you for your help.

Linda Degenstein
Director UCCRC
Transgenic/ES Cell Facility
The University of Chicago ldeg@midway.uchicago.edu




From YOCKEY at DCSMSERVER.MED.SC.EDU Mon Oct 21 19:15:02 1996
From: YOCKEY at DCSMSERVER.MED.SC.EDU (Courtland E. Yockey)

Subject: selection cassette-dependent phenotypes
Message-ID: <104196F7461@dcsmserver.med.sc.edu>

Hello,

I have a question regarding the phenotypes of targeted mutant mice.

One justification for flanking selection/mutation cassettes with loxP
sites is so that the cassette - and the transcription unit associated
with the selectable marker - can be removed in order to avoid an
artifactual mutant phenotype that is dependent upon its presence.

Does anyone know of published cases in which the phenotype of a mutant
mouse is influenced by the presence of the selectable marker itself
within the mutant locus? Cases in which the phenotype was compared,
for
instance, before and after Cre-mediated excision of a loxP-flanked
cassette?

Thanks for the information.

Courtland Yockey
-----------------------------------
Courtland Yockey, M.S. <yockey@dcsmserver.med.sc.edu>
from the lab of Noriko Shimizu, Ph.D.
                       <shimizu@dcsmserver.med.sc.edu>

University of South Carolina School of Medicine
Department of Developmental Biology & Anatomy
Columbia, SC 29208 USA

Phone# 803-733-1503 <lab>
Fax#   803-733-1533 <dpt>



From dolle at titus.u-strasbg.fr Tue Oct 22 15:19:24 1996
From: dolle at titus.u-strasbg.fr (Pascal DOLLE)

Subject: selection cassette-dependent phenotypes
Message-ID: <199610221213.OAA08832@titus.u-strasbg.fr >

>Hello,
>
>I have a question regarding the phenotypes of targeted mutant mice.
>
>One justification for flanking selection/mutation cassettes with loxP
>sites is so that the cassette - and the transcription unit associated
>with the selectable marker - can be removed in order to avoid an
>artifactual mutant phenotype that is dependent upon its presence.
>
>Does anyone know of published cases in which the phenotype of a mutant
>mouse is influenced by the presence of the selectable marker itself
>within the mutant locus? Cases in which the phenotype was compared,
for
>instance, before and after Cre-mediated excision of a loxP-flanked
>cassette?
>
>Thanks for the information.
>
>Courtland Yockey
>-----------------------------------

Dear Courtland,

a couple of years ago, we have reported that some 'weird' & dominant
phenotypes in chimeras harboring a Hoxd-10 disruption may have resulted
from integration effect of the neo cassette in the locus. At that time,
the
loxP technique was not developed, so we didn't have this mutation
without
the selection cassette. The ref is Rijli et al., Dev. Dynamics 201, p.
366
(1994).

I guess there will be more and more cases of mutations with or without
neo
cassettes coming out in the next future. You could contact Filippo
Rijli in
our institute who may have other examples of lines with or without TK-
neo
integrations.

Sincerely,



Pascal DOLLE, M.D., Ph.D.

CNRS, Unite 184 INSERM
I.G.B.M.C.
Parc d'innovation
B.P. 163
67404 ILLKIRCH Cedex
FRANCE

Tel: (33) 88 65 33 34 or 40
Fax: (33) 88 65 32 01
From dbowtell at petermac.unimelb.edu.au Wed Oct 23 09:50:38 1996
From: dbowtell at petermac.unimelb.edu.au (David Bowtell)

Subject: selection cassette-dependent phenotypes
In-Reply-To: <199610221213.OAA08832@titus.u-strasbg.fr >
Message-ID: <v03007800ae938c3f52ee@[203.4.164.101]>

As a follow on to the discussion about selectable markers we have
placed a
lacZneo fusion in frame downstream of the Sos1 gene. The marker is
driven
by the Sos1 promoter and there is no heterologous promoter sequences
inserted. We appear to see, however, mosaic expression of lacZ in some
heterozygous animals and wondered about silencing of the targeted locus
in
some tissues. Has anyone else encountered this?

******* Please note new email address **********
David Bowtell
Principal Research Fellow
Research Division
Peter MacCallum Cancer Institute
Locked Bag 1 A'Beckett St,
Melbourne 3000

Lab 61-3-96561287
Office 61-3-96561296
Fax 61-3-96561411
email: dbowtell@petermac.unimelb.edu.au




From anna at screams.gdb.org Wed Oct 23 10:29:45 1996
From: anna at screams.gdb.org (Anna Anagnostopoulos)

Subject: Mycoplasma Infection
Message-ID: <Pine.SOL.3.91.961023092228.11130A-100000@screams.gdb.org>


Hello everyone,

Dr. Yasuzawa is trying to rid his mice of a Mycoplasma infection.   I am
sure he would appreciate additional suggestions.

Thanks a lot

Anna V. Anagnostopoulos

> The reason why I noticed is that my mice are infected by mycoplasma.
> Especially, the clinical signs of old mice are more serious than
young.
> I'm worrying about both sudden death and expansion of contamination.
> I already start to feed tetracyclin to them. But, the condition has
not
> improved yet.
> If you give me any suggestion how to rescue them, I'm very happy.
>
>
> yours sincerely,
>
>   Kayoko Yasuzawa
>
>
>
>
>

~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Anna V. Anagnostopoulos, Ph.D.
Division of Biomedical Information Sciences
The Johns Hopkins University School of Medicine
2024 E. Monument Street
Baltimore, MD 21205-2236
tel: (410) 614-3226
fax: (410) 614-0434
e-mail: anna@gdb.org




From milstone at rascal.med.harvard.edu Wed Oct 23 12:26:12 1996
From: milstone at rascal.med.harvard.edu (David Milstone)

Subject: selection cassette-dependent phenotypes
Message-ID: <E0vG5BO-0005ZG-00@juliet.ic.ac.uk>

>Does anyone know of published cases in which the phenotype of a mutant
>mouse is influenced by the presence of the selectable marker itself
>within the mutant locus? Cases in which the phenotype was compared,
for
>instance, before and after Cre-mediated excision of a loxP-flanked
>cassette?
>>
>Courtland Yockey

Selectable markers have been   shown to influence the phenotype of mutant
mice by cis-mediated effects   on transciptional regulation (published
examples include beta globin   LCR element and CD11b). But this is
presumably
distinct from effects of the   coding region sequences of the selectable
marker itself.

David S. Milstone
Vascular Research Division
Department of Pathology
Brigham & Women's Hospital
LMRC 421
221 Longwood Avenue
Boston, MA
USA




From r-carver at nimr.mrc.ac.uk Thu Oct 24 17:40:21 1996
From: r-carver at nimr.mrc.ac.uk (Rick Carver)

Subject: Mycoplasma Infection
Message-ID: <13109.9610231552@nimsn01.nimr.mrc.ac.uk>

Dear All

>Dr. Yasuzawa is trying to rid his mice of a Mycoplasma infection. I
am
>sure he would appreciate additional suggestions.
>
>> The reason why I noticed is that my mice are infected by mycoplasma.
>> Especially, the clinical signs of old mice are more serious than
young.
>> I'm worrying about both sudden death and expansion of contamination.
>> I already start to feed tetracyclin to them. But, the condition has
not
>> improved yet.

My advice would be to rederive them, preferably by embryo transfer.
Antibiosis is a poor option because you can never be certain that it
has
been fully effective and you run a major risk of producing
tetracycline-resistant Mycoplasma. The NRC's 'Infectious Diseases of
Mice
and Rats' has a nice description of the protocol to be followed
post-caesarean rederivation. This protocol is equally valid if you
rederive
by embryo transfer. ET is much more microbiologically secure and if he
is
doing transgenic work he should already have the necessary equipment
and
skills available.

I hope that this is of assistance.




Rick Carver
Head of Biological Services and Institute Veterinarian
National Institute for Medical Research
The Ridgeway
MILL HILL
London                     E-Mail: r-carver@nimr.mrc.ac.uk
NW7 1AA                    Tel: + 44 (0)181 959 3666 ext 2199
United Kingdom             Fax: + 44 (0)181 913 8601
From dknudsen at scruznet.com Wed Oct 23 23:16:18 1996
From: dknudsen at scruznet.com (Dave Knudsen)

Subject: Mycoplasma Infection
Message-ID: <v01540b03ae94a16b55a4@[165.227.113.91]>

>Hello everyone,
>
>Dr. Yasuzawa is trying to rid his mice of a Mycoplasma infection. I
am
>sure he would appreciate additional suggestions.
>
>Thanks a lot
>
>Anna V. Anagnostopoulos
>
>> The reason why I noticed is that my mice are infected by mycoplasma.
>> Especially, the clinical signs of old mice are more serious than
young.
>> I'm worrying about both sudden death and expansion of contamination.
>> I already start to feed tetracyclin to them. But, the condition has
not
>> improved yet.
>> If you give me any suggestion how to rescue them, I'm very happy.

Anna -

I would agree with Rick Carver in that rederivation is the best way to
proceed for a contaminated colony. The assumption under this agreement
is
that the affected colony is a small breeding colony, and not a group of
stock animals that could be eradicated and replaced at the end of an
experiment. I also assume that your colleague, Dr. Yasuzawa, has
completed
the diagnostic process and confirmed mycoplasmosis as the problem. This
diagnosis can be made by serology, culture, or histopathology
(preferably
all three for a colony, to reduce the chance of false negative
findings).
The confirmation itself is important to resolution of the problem - I
can
think of several instances where other opportunistic pathogens,
environmental factors, immunodeficiencies, and/or transgenic phenotypes
have mimicked the clinical presentation of murine mycoplasmosis quite
closely. Also, I would add that antibiosis may help the clinical
appearance
of the animals by temporarily arresting disease progression, but it
will do
nothing in evoking a cure, and treated animals will continue to be
carriers
to infect new introductions (and births) into the colony.

I would personally be most comfortable, however, in dealing with the
rederivation issue by caesarian section; I suspect that confidence is
gained from the method most familiar. Over the years I have had more
positive experiences with caesarian rederivation in eliminating
pathogens
from mouse colonies, and embryo or ovarian manipulations seem always to
break at some point. Perhaps the best method could be determined by
close
questioning of the veterinary staff of the affected institution and
working
with them to resolve the outbreak.

Dave Knudsen DVM, DACLAM
Scotts Valley CA, USA
<dknudsen@scruznet.com)




From pasceri at sickkids.on.ca Tue Oct 1 21:34:58 1996
From: pasceri at sickkids.on.ca (Peter Pasceri)

Subject: Cryopreservation
Message-ID: <v01510100ae772f68646f@[142.20.24.42]>

Hi all!

I am looking for suppliers of mouse embryo freezing kits. Can you help?

Thanks,

Peter

--
Peter Pasceri
pasceri@sickkids.on.ca
Genetics Research                                        Phone:     (416)
813-4205
The Hospital for Sick Children                           Toronto,
Ontario,Canada




From p.sobieszczuk at ic.ac.uk Thu Oct 3 11:37:29 1996
From: p.sobieszczuk at ic.ac.uk (Peter Sobieszczuk)

Subject: Transgenic mice/Transplantation (fwd)
Message-ID: <v01510104ae7954f36393@[155.198.45.54]>

Sorry, if it is a duplication for some of you.
Peter (your listowner)


>Hello everyone,
>
>Any advise/suggestions on the following questions would be greatly
>appreciated.
>
>Thank you
>
>Anna V. Anagnostopoulos
>TBASE
>Division of Biomedical Information Sciences
>The Johns Hopkins University School of Medicine
>anna@gdb.org
>
>---------------
>
>I am working with mice, doing Bone Marrow and Peripheral Blood
>Tranplantation experiments, at the Department of Immunogenetics of the
New
>York Blood Center. We are interested in developing a syngeneic mouse
>transplantation model. We would like to use as donors, animals
carrying a
>transgene, that its product is not lethal or significantly affecting
the
>recipients (especially hematologically), and also is easily detectable
>post-transplant. For example, mice transgenic for a human gene,
encoding
>for a human antibody, would be a good model. The idea would be to show
>that the donor cells (positive for the transgene) engraft, and the
>transgene is functional (we can detect the product) after transplant.
The
>mouse strains I am working with at the present time are C57BL/6, A/J
and
>B6A (C57BL/6 X A/J) (all obtained from Jackson Labs). It would help
>tremendously if the transgenic animals are of the same background. Do
you
>have any suggestions?
>




From mangia at uniroma1.it Fri Oct 4 12:46:36 1996
From: mangia at uniroma1.it (Franco Mangia)

Subject: Transgenic mice/Transplantation (fwd)
Message-ID: <E0v98iC-0006aA-00@juliet.ic.ac.uk>


>
>We are interested in developing a syngeneic mouse
>>transplantation model. We would like to use as donors, animals
carrying a
>>transgene, that its product is not lethal or significantly affecting
the
>>recipients (especially hematologically), and also is easily
detectable
>>post-transplant. For example, mice transgenic for a human gene,
encoding
>>for a human antibody, would be a good model. The idea would be to
show
>>that the donor cells (positive for the transgene) engraft, and the
>>transgene is functional (we can detect the product) after transplant.
The
>>mouse strains I am working with at the present time are C57BL/6, A/J
and
>>B6A (C57BL/6 X A/J) (all obtained from Jackson Labs). It would help
>>tremendously if the transgenic animals are of the same background. Do
you
>>have any suggestions?
>>
>Anna V. Anagnostopoulos
>TBASE
>Division of Biomedical Information Sciences
>The Johns Hopkins University School of Medicine
>anna@gdb.org



I think that the ROSA26 transgenic mice, developed by Phil Soriano, are
just what you need for labeling engrafts at a single cell level. These
mice carry a lacZ transgene directed by a ubiquitous promoter active in
all
tissues, without apparently affecting cell viability, since these mice
are
fine. These mice can be obtained from the Jackson Labs either as
B6,129 F1
(the original Soriano's hybrid strain) or with a pure C57BL/6J
background.
Complete names of these mice are: "B6,129-TgR(ROSA26)26 Sor", and
"C57Bl/6J-TgR(ROSA26)26 Sor".   I am also very interested in obtaining
an
ES cell line(s) from these mice. Is anybody aware whether these ES
lines
are being developed in some lab? Also, does anybody know whether
ROSA26
mice with nuclearly targeted beta-gal have been produced?
Franco



Franco Mangia
Laboratory of General Biology
Department of Psychology
University La Sapienza of Rome
c/o Institute of Histology and General Embryology
Via Borelli, 50
00161 Rome, Italy
tel: 39-6-4976-8103   FAX: 39-6-4976-8099
E-mail: mangia@uniroma1.it




From p.sobieszczuk at ic.ac.uk   Sat Oct   5 18:19:10 1996
From: p.sobieszczuk at ic.ac.uk (Peter Sobieszczuk)

Subject: "TgList Backstage" (admin. news *2) ARCHIVES & TBASE
Message-ID: <v01510102ae7c443e5426@[155.198.45.54]>

Dear Transgenic Subscribers,

Greetings to newcomers,

ARCHIVES
Charmaine Foltz asked me some time ago:
>I was wondering if the information on the list will be retrievable
>(i.e., will there be a data base that we can search archived files).

very much so, send the message:
                index transgenic-list

to listserver at
                   majordomo@ic.ac.uk

and you shall receive an index of monthly files, something like:

>>>> index transgenic-list
total 241
-rw-rw---- 1 daemon       10364    Jul 29 16:38 transgenic-
list.archive.9607
-rw-rw---- 1 majordom    146666    Aug 29 21:36 transgenic-
list.archive.9608
-rw-rw---- 1 majordom     71873    Sep 25 15:31 transgenic-
list.archive.9609
-rw-rw---- 1 majordom      6715    Oct   4 12:46 transgenic-
list.archive.9610
>>>>

where July entries file name is "transgenic-list.archive.9607"

If you then send:
        get transgenic-list transgenic-list.archive.9607

you should receive all postings for that month (in this case, July) as
individual e-mail messages.

If you have access to world wide web, (WWW) you can explore archives
with
the luxury of selecting individual entries etc., using your favourite
browser, Netscape, Mosaic or Explorer.

The URL address is:
        http://www.lists.ic.ac.uk/hypermail/transgenic-list

I'm afraid however that even with the browser, files are not searchable
(at
this stage anyway), although subject, (which should be the topic of any
message) is clearly visible.


TBASE
Anna Anagnostopoulos who is co-ordinating TBASE data base site sent
this
introduction (with minor editorial changes P.S.):

-----------------------------------------------------------------------
-
BEGINNING OF TEXT

TBASE, the Transgenic/Targeted Mutation Database, is an attempt to
organize information on transgenic animals and targeted mutations
generated, and analyzed worldwide. TBASE is available on the WWW

http://www.gdb.org/Dan/tbase/tbase.html

and can be searched either
quite broadly across the entire database (the 'blunt object' approach)
or
quite specifically with the use of 28 distinct searchable fields (the
'scalpel and tweezers' approach). TBASE contains a wealth of
information
about each animal in the database. A selection of the fields present in
the databases are: line name, genus, DNA construct description, host
background, line source, method used , transgene/targeted allele
expression, lethality, phenotype, handling, contact information, full
citation and abstracts of the papers related to the creation of a line.

Owing to the wide use of homologous recombination in ES cells,
TBASE initially focused on maintaining a comprehensive coverage of
all targeted mutations reported to date. Moreover, it focused on the
mouse
as the predominant mammalian model. Specific emphasis is still placed
on
cataloging novel knockout mice, which have defined genetic
modifications,
with an extensive detailed description of their resulting mutant
phenotypes. In the future, the focus of TBASE will expand to cover
all animals resulting from transgenesis or targeted mutation,
regardless
of species. Additionally, TBASE plans to incorporate 'transgenic
knockouts', that is, knockout animals that have subsequently served as
recipients for transgenes.

TBASE should be viewed as a dynamic database with a potentially
unlimited
number of entries, expected to grow considerably as more experimental
methods and lines are generated. Data acquisition is primarily effected
by
active literature scanning and manual data entry, as well as processing
of
direct submission forms.

Literature scanning refers to direct data extraction from the
scientific
literature through regular examination of over 20 journals. These
journals have been statistically identified as periodically stable
sources
of information on transgenesis and gene targeting. Active literature
scanning has, so far, been the primary mode of data accumulation, and
has
ensured that the database faithfully reflects the general direction of
related research. It is expected to remain a significant component of
the
data acquisition tactics, even as other methods of data acquisition
become
available (see below). In an effort to avoid missing important data,
keyword-related references from additional journals are also screened
by
the use of Medline and Current Contents.

In the future, data that are not manually entered by TBASE staff
through the literature scanning process, are expected to arrive either
as
direct paper or as electronic submissions. Although over 98% of the
data
have been entered manually so far, direct paper submissions are
expected
to increase as TBASE becomes fully adopted by the scientific
community. In addition, as electronic submission tools become
available,
more research groups will be prepared to commit resources to the
production of data deposited into TBASE. Electronic submission
tools and their error-checking capabilities will ensure efficient and
timely deposition of the data.

END OF TEXT
-----------------------------------------------------------------------
-----


STATISTIC:
There are approx. 240 members from at least 18 countries registered
now,
and I'm starting to think aren't we growing too big, read to much to
read.

Regards,

Yours sincerely,

Listowner




_______________________________________________________________________
____

Dr Peter Sobieszczuk
Imperial College School of Medicine at St. Mary's        tel: 0171 594
3784
Biochemistry and Molecular Genetics                      tel: 0171 594
3799
Norfolk Place                                            fax: 0171 706
3272
London W2 1PG, U.K.                          e-mail:
p.sobieszczuk@ic.ac.uk
_______________________________________________________________________
____




From khelmuth at facstaff.wisc.edu Wed Oct 9 15:28:47 1996
From: khelmuth at facstaff.wisc.edu (khelmuth@facstaff.wisc.edu)

Subject: fire codes within animal colony
Message-ID: <v02140b00ae7ef3eb32c0@[144.92.94.130]>

Hello everyone-

I face a delemma within our facility and am hoping someone can help me
out.
We recently moved our rat and mouse colonies into a new building.
Everything seems to be going well except for one thing. We experienced
the
first fire drill and realized that there are 3 fire alarms within our
5ft.
wide clean hallway and 3 fire alarms within our 5ft. wide dirty
hallway.
They are extremely loud and potentially problematic for our animals.
Our
building manager agrees that this seems extremely overdone and wishes
to
get some of the alarms diconnected. However, the state fire code
regulator
feels that with the background noises of the cage washer, movement of
carts
and racks, etc. that these extra fire alarms are necessary. He will
only
follow suit if I can come up with evidence that not only does it affect
the
well-being of the animals but that other facilities have a small number
of
alarms. I have obtained documentation of loud noises and it's effect
on
breeding, embryo production, etc. Can someone help me out with
regulations
regarding fire alarms within your animal facility?

Any suggestion on how to deal with this would be beneficial.
Thanks

Kathy Krentz Helmuth

Kathy Krentz Helmuth
Transgenic Facility
425 Henry Mall
Room 2210 Biotechnology Center
Madison, WI 53706
(608) 265-2801
email:khelmuth@facstaff.wisc.edu




From landel at medsci.udel.edu Wed Oct 9 15:44:03 1996
From: landel at medsci.udel.edu (Carlisle Landel)

Subject: fire codes within animal colony
Message-ID: <199610091444.KAA19359@helios.medsci.udel.edu>

I'm curious--how often do these alarms go off? Maybe instead you could
get them to absolve you from frequent fire drills.

Carlisle Landel

>From owner-transgenic-list@ic.ac.uk Wed Oct 9 10:35:44 1996
>Mime-Version: 1.0
>Content-Type: text/plain; charset="us-ascii"
>To: transgenic-list@ic.ac.uk
>From: khelmuth@facstaff.wisc.edu
>Subject: fire codes within animal colony
>Sender: owner-transgenic-list@ic.ac.uk
>Precedence: bulk
>Reply-To: transgenic-list@ic.ac.uk
>
>Hello everyone-
>
>I face a delemma within our facility and am hoping someone can help me
out.
>We recently moved our rat and mouse colonies into a new building.
>Everything seems to be going well except for one thing. We
experienced the
>first fire drill and realized that there are 3 fire alarms within our
5ft.
>wide clean hallway and 3 fire alarms within our 5ft. wide dirty
hallway.
>They are extremely loud and potentially problematic for our animals.
Our
>building manager agrees that this seems extremely overdone and wishes
to
>get some of the alarms diconnected. However, the state fire code
regulator
>feels that with the background noises of the cage washer, movement of
carts
>and racks, etc. that these extra fire alarms are necessary. He will
only
>follow suit if I can come up with evidence that not only does it
affect the
>well-being of the animals but that other facilities have a small
number of
>alarms. I have obtained documentation of loud noises and it's effect
on
>breeding, embryo production, etc. Can someone help me out with
regulations
>regarding fire alarms within your animal facility?
>
>Any suggestion on how to deal with this would be beneficial.
>Thanks
>
>Kathy Krentz Helmuth
>
>Kathy Krentz Helmuth
>Transgenic Facility
>425 Henry Mall
>Room 2210 Biotechnology Center
>Madison, WI 53706
>(608) 265-2801
>email:khelmuth@facstaff.wisc.edu
>
>
>
>
>



From duffyh at war.wyeth.com Wed Oct 9 19:45:06 1996
From: duffyh at war.wyeth.com (Heidi Duffy)

Subject: fire codes within animal colony -Reply
Message-ID: <s25bb9fa.076@war.wyeth.com>

I used to work in animal facility that used flashing lights instead of
 audible alarms within the animal facility. The audible alarms were
only in the office areas of the facility. (Our phones within the
facility were set up the same way, including the cagewash areas).
Designated techs were assigned to check all animal rooms for
personnel evacuation before leaving the buildings. It worked out very
well.

Heidi Duffy
email:duffyh@war.wyeth.com




From david_gask at Merck.Com Thu Oct 10 13:47:00 1996
From: david_gask at Merck.Com (David Gask)

Subject: Fire Alarm problems-Kathy Krentz Helmuth
Message-ID: <199610101150.HAA21899@igw2>


Kathy,
              Why don't you investigate the use of ' SILENTONE ALARMS'
as
an alternative. These are used widely in the UK and were developed to
prevent problems for the animals caused by more conventional sounders.
They
produce a loud noise suitable for use as an alarm in the event of fire,
but
at the same time the frequency is below the threshold frequency, or
outside
the most sensitive range for most commonly used laboratory animals. If
you
require the address of a supplier I can send it to you. All our animal
facility fire alarms are of this type.

Hope this helps

Dave Gask
Operations Coordinator
Merck Sharp & Dohme
Neuroscience Research Centre
Terlings Park
Eastwick Road
Harlow, Essex
CM20 2QR
England
Tel 01279 440220
e-mail David_Gask@Merck.com


Hello everyone-

I face a delemma within our facility and am hoping someone can help me
out.
We recently moved our rat and mouse colonies into a new building.
Everything seems to be going well except for one thing. We experienced
the
first fire drill and realized that there are 3 fire alarms within our
5ft.
wide clean hallway and 3 fire alarms within our 5ft. wide dirty
hallway.
They are extremely loud and potentially problematic for our animals.
Our
building manager agrees that this seems extremely overdone and wishes
to
get some of the alarms diconnected. However, the state fire code
regulator
feels that with the background noises of the cage washer, movement of
carts
and racks, etc. that these extra fire alarms are necessary. He will
only
follow suit if I can come up with evidence that not only does it affect
the
well-being of the animals but that other facilities have a small number
of
alarms. I have obtained documentation of loud noises and it's effect
on
breeding, embryo production, etc. Can someone help me out with
regulations
regarding fire alarms within your animal facility?
Any suggestion on how to deal with this would be beneficial.
Thanks

Kathy Krentz Helmuth



From Roger.Leemann at imr.psi.ch Thu Oct 10 14:21:53 1996
From: Roger.Leemann at imr.psi.ch (Roger C. Leemann)

Subject: Rodent-research list
Message-ID: <v01540b00ae82a600b461@[129.129.90.92]>

Hi

Since I'm working with mice but do not use transgenic animals, I
subscribed
to the rodent-research list about a month ago. Majordomo told me, that
it
had forwarded my request to the list owner for approval but I haven't
got
any response since.
Maybe it's not a list for every Jack and Joe (or was it Jack and
Jill?), or
maybe the list owner is too busy, or on vacation, whatever. I don't
mind if
they don't want me, but ones likes to know...
Anybody in this list who has a clue why the rodent-research list seems
dead?

Roger

:::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::
:::::
Work: Institute for Medical Radiobiology        ++41 (56) 310-3792
[desk]
       CH-5232 Villigen PSI                     ++41 (1) 3856-561
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       Switzerland                              ++41 (56) 310 3294
[fax]
Home: Nordstrasse 26, CH-8006 Zuerich
       Switzerland                              ++41 (1) 361 0349
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From p.sobieszczuk at ic.ac.uk Thu Oct 10 16:55:30 1996
From: p.sobieszczuk at ic.ac.uk (Peter Sobieszczuk)

Subject: Rodent-research list & "TgL Backstage"(admin.news*3)
Message-ID: <v01510100ae82cd2a148c@[155.198.45.54]>

Dear Roger,
>Since I'm working with mice but do not use transgenic animals, I
subscribed
>to the rodent-research list about a month ago. Majordomo told me, that
it
I presume listserver "majordomo" at Caltech, and not "ic.ac.uk" at
Imperial College?

>Maybe it's not a list for every Jack and Joe (or was it Jack and
Jill?), or
Do not take it personnally, it's just a computer.

>maybe the list owner is too busy, or on vacation, whatever. I don't
mind if
>they don't want me, but ones likes to know...

Sorry, I still do not know what exactly happened, Eric Mercer had some
technical problems supporting "rodent-research" list, 3 or 4 months
ago. A few people has tried to contact him, at the end, this list was
created (see the introduction to the list available also by sending
"info transgenic-list" message to "majordomo@ic.ac.uk" address).

You have been with us (transgenic-list) for the month now and probably
noticed that the list is un-moderated and has no restriction on the
content. Transgenic - is just a name on a server, and the list a quick
way to exchange information.

I would not like to start a discussion on definition of transgenesis,
but would imagine that for the active researchers in the field it would
cover quite a bit of biology, from molecular to developmental, with the
animal models not exclussively restricted to rodents.

So it is not the name but participants who will take the discussion in
any particular direction, including yourself.

Who would you like to see on the list Roger, or should I ask everybody:

Who else do you think we should invite to the list?

Best regards,

Sincerely,

Peter - your listowner




From s.tan at anatomy.unimelb.edu.au Tue Oct 15 05:09:53 1996
From: s.tan at anatomy.unimelb.edu.au (S.Tan)

Subject: Fostering baby mice
Message-ID: <v03007812ae88bef7a8eb@[128.250.225.71]>
I am grafting pieces of neural tissue into the cortex of newborn mice
but
find that the mother keeps eating the babies. The wound is closed with
a
single silk suture and this appears to attract the attention of mum.
The
mother's strain is C57/BL6 x DBA. Are there less aggressive strains of
mums out there? Thanks.




From a.annala at ucl.ac.uk Tue Oct 15 06:51:24 1996
From: a.annala at ucl.ac.uk (Dr. Alexander J. Annala)

Subject: Fostering baby mice
Message-ID: <v01540b00ae88e518e7ca@[128.40.81.204]>

>I am grafting pieces of neural tissue into the cortex of newborn mice
but
>find that the mother keeps eating the babies. The wound is closed
with a
>single silk suture and this appears to attract the attention of mum.
The
>mother's strain is C57/BL6 x DBA. Are there less aggressive strains
of
>mums out there? Thanks.

Most F1's make very good mothers. Your C57BL/6 x DBA are F1 mice.
We use C57BL/6 x CBA and C57BL/10 x CBA with good results.

Do you take precaution to avoid leaving scent on operated baby mice?
Some people roll babies in mom's urine before returning them to the
nest. Others treat mom's nose with alcohol to keep her from smelling
any differences between babies for awhile. This might be a scent issue
rather than an exposed suture problem.

Internalize suture might be difficult in baby mice. Maybe you could
make your incision somewhere away from your skull opening, slide
the skin opening over the area you want to open bone, do your work,
slide the skin back, and maybe use one of the tissue adhesives which
are pretty available around medical schools and operating theatres
to close the wound. This would avoid leaving exposed suture for the
mom to chew on.




From fobo at ltk.unizh.ch Tue Oct 15 07:22:40 1996
From: fobo at ltk.unizh.ch (Frank Bootz)

Subject: Fostering baby mice
Message-ID: <1.5.4.32.19961015062240.0068aa74@rzu-mailhost.unizh.ch>

Dear colleaque,
I have done thymectomy in newborn mouse. The wound was also closed with
a
single U-suture in the neck. The pups awake within 5 minutes after
methofane
(methoxyflurane) anesthesia and brought back to the mother immediately.
We
don't anesthetize the mother, contrary the most papers describing this
procedure. In this case the mother's strain is C57/Bl/6 x PO. If we can
choose the strain, we normally use Zur:ICR original Charles River swiss
mice
CD-1. They are mice with excellent mother attributes.

Best regards,

Frank
Frank Bootz
Institute of Laboratory Animal Science
University of Zurich


Winterthurerstrasse 190                     phone: +41 1 257 54 55
8057 Zurich                                 fax:   +41 1 257 57 03
Switzerland                                 e-mail fobo@ltk.unizh.ch




From GWOLFF at NCTR.FDA.GOV Tue Oct 15 14:44:51 1996
From: GWOLFF at NCTR.FDA.GOV (George L. Wolff)

Subject: Fostering baby mice
Message-ID: <695B452EEE@EFS.NCTR.FDA.GOV>

Zur:ICR cannot be descended from CD-1 mice - or the strain
designation is incorrect. Let me know if you'd like more details.
George L. Wolff



Tel: (501) 543-7522
FAX: (501) 543-7635/7662
NCTR       HFT-140
3900 NCTR Road
Jefferson, AR 72079-9502




From gotto at leland.Stanford.EDU Wed Oct 16 20:03:00 1996
From: gotto at leland.Stanford.EDU (Glen Otto)

Subject: Fostering baby mice
In-Reply-To: <1.5.4.32.19961015062240.0068aa74@rzu-mailhost.unizh.ch>
Message-ID: <v03007800ae8adbf9a0a6@[171.65.18.36]>
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Type: text/enriched
Size: 577 bytes
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From a.annala at ucl.ac.uk Thu Oct 17 05:50:59 1996
From: a.annala at ucl.ac.uk (Dr. Alexander J. Annala)

Subject: Fostering baby mice -- Failure to thrive?
Message-ID: <E0vDcxZ-0002Gs-00@juliet.ic.ac.uk>

What can be done to assist baby mice in distress after birth? Some
chimeras with neuroreceptor channel mutations experience difficulty
feeding themselves -- and sometimes even suckling milk from mother.

We have in the past injected approx 20% weight of animal with Ringers
Lactate (Hartman's Solution). However there must be more aggressive
therapy which would facilitate survival of distressed baby mice.

We have also used incubator -- with reservations about the possibility
of dehydration from elevated environmental temperature.

Thanks,

Alexander J. Annala, Ph.D.
Senior Research Fellow
Laboratory for Molecular Pharmacology
University College London




From carton at murray.fordham.edu Wed Oct 16 23:44:40 1996
From: carton at murray.fordham.edu (Jill Carton)

Subject: knockout mouse construct problems
Message-ID: <v02130500ae8b106fe16f@[169.132.99.88]>

        We are working on generating several knockout mice and have
been
successful in producing transgenes for some of our genes, but we have a
fairly consistent problem with the final ligation step of assembling
the
construct. When we PCR our ligation reaction to see if the DNA pieces
have
ligated we do get the appropriate product however analysis of the
clones
by miniprep show no positive clones. So we are suspecting that the
problem
is in the uptake or growth of the construct in the bacterial cells.
Although, each fragment of the transgene can be propagated in the
bacteria.
        At this stage we are working with a very large piece of DNA
(~12KB)- and perhaps this will lower the ligation frequency-but we have
generated larger plasmids with little problem.
        We are using JM109 or DH5a as our competent cells (chemically
competent). Any suggestions at all would be greatly appreciated!

Jill Carton
Dept. Biological Sciences
Fordham University
Larkin Hall, rm 160
Bronx, NY 10458
718-817-3652
718-817-3645 (fax)
carton@murray.fordham.edu




From p.sobieszczuk at ic.ac.uk Thu Oct 17 15:23:53 1996
From: p.sobieszczuk at ic.ac.uk (Peter Sobieszczuk)

Subject: Nod/Scid mice
Message-ID: <v01510103ae8bffb476b7@[155.198.45.54]>

Forwarded from [Anna Anagnostopoulos <anna@screams.gdb.org>]


>Hello everyone,
>
>Dr. Sally J. Cutler (Department of Medical Microbiology, Charing Cross
>Hospital, London. Telephone:+ 44181 846 7570) is looking for sources
of
>Nod/Scid mice, preferably in the UK. Any suggestions on where to
look?
>
>Thank so much
>
>~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
>Anna V. Anagnostopoulos, Ph.D.
>Division of Biomedical Information Sciences
>The Johns Hopkins University School of Medicine
>2024 E. Monument Street
>Baltimore, MD 21205-2236
>tel: (410) 614-3226
>fax: (410) 614-0434
>e-mail: anna@gdb.org




From james at icr.ac.uk Thu Oct 17 22:00:12 1996
From: james at icr.ac.uk (james wallace)

Subject: Nod/Scid mice
Message-ID: <SIMEON.9610171712.A@sbp112.icr.ac.uk>
>

> >Dr. Sally J. Cutler (Department of Medical Microbiology, Charing
Cross
> >Hospital, London. Telephone:+ 44181 846 7570) is looking for
sources of
> >Nod/Scid mice, preferably in the UK. Any suggestions on where to
look?
> >
> >Thank so much
> >
> >~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
> >Anna V. Anagnostopoulos, Ph.D.
> >Division of Biomedical Information Sciences
> >The Johns Hopkins University School of Medicine
> >2024 E. Monument Street
> >Baltimore, MD 21205-2236
> >tel: (410) 614-3226
> >fax: (410) 614-0434
> >e-mail: anna@gdb.org
>
> Hi,
        Tell Sally to give me a call as we have a colony
of Nod\Scids here at ICR in the UK.

Best Wishes


Jim Wallace
Institute of Cancer Research
The McElwain Laboratories
15 Cotswold Rd
Belmont Sutton
Surrey SM2 5NG
England
tel:    0181 643 8901 Ext 4638
Fax:    0181 770 1395
e_mail james@icr.ac.uk
>
>




From anna at screams.gdb.org Thu Oct 17 18:18:07 1996
From: anna at screams.gdb.org (Anna Anagnostopoulos)

Subject: Available enhancer traps
Message-ID: <Pine.SOL.3.91.961017131414.2600G-100000@screams.gdb.org>


Hello again,

If any of you is interested in Joe Miano's enhancer traps or knows of a
bulletin where he can have the pictures posted please let me know.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Anna V. Anagnostopoulos, Ph.D.
Division of Biomedical Information Sciences
The Johns Hopkins University School of Medicine
2024 E. Monument Street
Baltimore, MD 21205-2236
tel: (410) 614-3226
fax: (410) 614-0434
e-mail: anna@gdb.org


---------- Forwarded message ----------
Date: Wed, 16 Oct 1996 16:36:44 -0500
From: Joe Miano <jmiano@post.its.mcw.edu>
To: anna@gdb.org
Subject: enhancer trap


We do transgenics and I have two questions:

(1) Would you or anyone you know be interested in an enhancer trap
mouse
that has expression in a discrete region of each limb as well as the
face?
I hesitate to kill these mice off because I can't help but think
someone,
somewhere might be interested.

(2) Do you know of a bulletin board for posting pictures of enhancer
trap
(gene trap etc) mice? I think such a board would be quite valuable.
For
example, we have genrated several enhancer trap mice (one that stained
a
region of the brain and the spinal cord is regrettably gone). None of
these
mice are of interest to us since we are looking for a SMC-restricted
pattern
of expression (see SM22 transgenic mouse paper in March '96 issue of
JCB).
There may be others, however, who would gladly accept such mice.


Sincerely,

Joseph M. Miano, Ph.D.




From kelly at citi2.fr Fri Oct 18 05:42:58 1996
From: kelly at citi2.fr (U344)

Subject: knockout mouse construct problems
Message-ID: <v02120d03ae8cb9b6d37f@[194.254.89.19]>

>        At this stage we are working with a very large piece of DNA
>(~12KB)- and perhaps this will lower the ligation frequency-but we
have
>generated larger plasmids with little problem.
>        We are using JM109 or DH5a as our competent cells (chemically
>competent). Any suggestions at all would be greatly appreciated!

        Whenever I'm forced to deal with DNA of such a size, I use the
Hanahan transformation proceedure to produce competent cells... the
original reference is J. Mol. Biol. 1983, (166) pg 557... it may also
be
to your advantage to lift and screen colonies before picking them.

                                good luck...

INSERM Unite 344 Endocrinologie Moleculaire
156 rue de Vaugirard, 75730 Paris cedex 15 France

'Les raisonables ont dure, les passiones ont vecu'




From ldeg at midway.uchicago.edu Fri Oct 18 23:19:37 1996
From: ldeg at midway.uchicago.edu (Linda Degenstein)

Subject: Transgenic Facility Yellow Pages
Message-ID: <199610182219.RAA15525@midway.uchicago.edu>

    I am often contacted by people outside my university inquiring as to
whether or not we can produce KO or transgenic mice for them. These are
often from investigators out of the state or even out of the country. I
try
to match them up with an institution in their location if I know of
one.

   I would like to make a list of all the known facilities, with
pertinent
info, such as institution, location, services offered, contact person,
telephone and e-mail listings, and whether or not you can perform these
services for someone outside your institution, but in your area.

   If you are willing to e-mail this info, I will summarize in about a
week
for both lists.

   Thank you for your help.

Linda Degenstein
Director UCCRC
Transgenic/ES Cell Facility
The University of Chicago
ldeg@midway.uchicago.edu




From pinkert at cmed.bhs.uab.edu Fri Oct 18 18:31:33 1996
From: pinkert at cmed.bhs.uab.edu (Dr. C.A. Pinkert)

Subject: Transgenic Facility Yellow Pages
Message-ID: <19961018225449159.AAA83@compaq.uab.edu>

>   Date:        Fri, 18 Oct 1996 17:19:37 -0500 (CDT)
>   To:          transgenic-list@ic.ac.uk, compmed@WUVMD.WUSTL.edu
>   From:        ldeg@midway.uchicago.edu (Linda Degenstein)
>   Subject:     Transgenic Facility Yellow Pages
>   Reply-to:    transgenic-list@ic.ac.uk

>     I am often contacted by people outside my university inquiring as
to
> whether or not we can produce KO or transgenic mice for them. These
are
> often from investigators out of the state or even out of the country.
I try
> to match them up with an institution in their location if I know of
one.
>
>     I would like to make a list of all the known facilities, with
pertinent
> info, such as institution, location, services offered, contact
person,
> telephone and e-mail listings
I think you have info for the NICHD facility at UAB. I'm not sure if
such a listing of all facilities will be problematic with the patent
positions that DNX
(as well as GenPharm, Lexicon, etc. for ES cell work) are staking
out. But so long as the caution is there in advance, it may all be OK.

Carl



From a.annala at ucl.ac.uk Sun Oct 20 14:32:43 1996
From: a.annala at ucl.ac.uk (Dr. Alexander J. Annala)

Subject: Transgenic Facility Yellow Pages
Message-ID: <v01540b00ae8fe7874a90@[128.40.81.204]>

Dr. Pinkert,

I am curious about the patent claims you mention. We do some
pronuclear
microinjection of DNA, ES cell injection into blastocysts, and
aggregation
of ES cells with morula to produce transgenic mice. Are individuals
and/or
organizations claiming patent rights which would preclude carrying out
these basic transgenic procedures in academic laboratories? Are
license
fees to obtain the right to perform these procedures anywhere within
the
reach of small academic laboratories?

Are the basic transgenic patent claims analogous to the original
Stanford
recombinant DNA claims where a $10,000 fee was required for any lab
to perform basic molecular biology techniques? It would seem few, if
any, academic laboratories pay this fee today.

It might be interesting for people to have an up to date listing
available on
the network containing a summary of transgenic relevant patent claims
and licensing arrangements/fees.

Thanks,

Alexander J. Annala, Ph.D.
Wellcome Senior Research Fellow
Laboratory for Molecular Pharmacology
University College London
Gower Street
London WC1E 6BT

Tel: +44(171)380-7857
Fax: +44(171)380-7245
Email: a.annala@ucl.ac.uk

>> Date:           Fri, 18 Oct 1996 17:19:37 -0500 (CDT)
>> To:             transgenic-list@ic.ac.uk, compmed@WUVMD.WUSTL.edu
>> From:           ldeg@midway.uchicago.edu (Linda Degenstein)
>> Subject:        Transgenic Facility Yellow Pages
>> Reply-to:       transgenic-list@ic.ac.uk
>
>>     I am often contacted by people outside my university inquiring as
to
>> whether or not we can produce KO or transgenic mice for them. These
are
>> often from investigators out of the state or even out of the
country. I try
>> to match them up with an institution in their location if I know of
one.
>>
>>     I would like to make a list of all the known facilities, with
pertinent
>> info, such as institution, location, services offered, contact
person,
>> telephone and e-mail listings
>I think you have info for the NICHD facility at UAB. I'm not sure if
>such a listing of all facilities will be problematic with the patent
>positions that DNX
>(as well as GenPharm, Lexicon, etc. for ES cell work) are staking
>out. But so long as the caution is there in advance, it may all be
OK.
>
>Carl




From pinkert at cmed.bhs.uab.edu Sun Oct 20 10:02:00 1996
From: pinkert at cmed.bhs.uab.edu (Dr. C.A. Pinkert)

Subject: Transgenic Facility Yellow Pages
Message-ID: <19961020142523272.AAA152@compaq.uab.edu>

> Dr. Pinkert,
>
> I am curious about the patent claims you mention. We do some
pronuclear
> microinjection of DNA, ES cell injection into blastocysts, and
aggregation
> of ES cells with morula to produce transgenic mice. Are individuals
and/or
> organizations claiming patent rights which would preclude carrying
out
> these basic transgenic procedures in academic laboratories?
If you are providing services in house, there should be no problem.
If you provide services outside your home institution, you can be
liable to the folks holding patents on all the microinjection/gene
transfer patents that have been awarded.
  Are license
> fees to obtain the right to perform these procedures anywhere within
the
> reach of small academic laboratories?
When it is allowed by the patent holders. But if the work is
in-house AND for non-commercial basic research - most of the
companies with the rights will leave you alone. It is only when you
offer a service or begin toward commercialization that you will find
yourself in need of appropriate legal counsel.
>
> Are the basic transgenic patent claims analogous to the original
Stanford
> recombinant DNA claims where a $10,000 fee was required for any lab
> to perform basic molecular biology techniques? It would seem few, if
> any, academic laboratories pay this fee today.
Agreed, but as an example, for DNA microinjection outside of your
home institution, DNX would not allow you to provide the service (as
it competed with their program - but in-house basic research was
automatically exempted). Now, if you want to commercialize or
transfer animals, they have a basic fee structure in mind. However,
if you agree to provide microinjection services to say another
institution - even at/below your actual costs, you would likely be
liable for treble damages . . .
>
> It might be interesting for people to have an up to date listing
available on
> the network containing a summary of transgenic relevant patent claims
> and licensing arrangements/fees.
I agree, but some of the ES cell patents as well as more recent
targeting/expression patents are in process and disclosure is
not readily available.
>
Good luck.

Carl A. Pinkert



From stewarv at cesmtp.ccf.org Mon Oct 21 16:27:16 1996
From: stewarv at cesmtp.ccf.org (Valerie Stewart MS)

Subject: Transgenic Facility Yellow Pages -Reply
Message-ID: <s26b5f4a.015@cesmtp.ccf.org>

Hi Linda--

Good idea, though with the patent battles, our reach is limited. Our
policy so far has been to make mice for Clinic investigators and their
collaborators. Here is the information you requested:

Transgenic/IKnockout Core Facility
Cleveland Clinic Research Institute
FF6-50, 9500 Euclid Avenue
Cleveland, OH 44106
Director, Valerie Stewart
Technician, Christine Brant
phone (216)444-8521
fax (216)445-6257
e-mail "stewarv@cesmtp.ccf.org"
Services include pronuclear injection, blastocyst injection,
cryopreservation by vitrification, training in all techniques. We
also do microinjection of proteins, etc. into cell lines via an
Eppendorf automated system.
Services and prices can also be accessed through the Clinic's web
page at "www.ccf.org". Go to Main Menu, then Research Institute,
then Scientific Support Services, then Core Services.

>>> Linda Degenstein <ldeg@midway.uchicago.edu> - 10/18/96 6:19 PM
>>>
    I am often contacted by people outside my university inquiring as
to whether or not we can produce KO or transgenic mice for them.
These are often from investigators out of the state or even out of
the country. I try to match them up with an institution in their
location if I know of one.

   I would like to make a list of all the known facilities, with
pertinent info, such as institution, location, services offered,
contact person, telephone and e-mail listings, and whether or not you
can perform these services for someone outside your institution, but
in your area.

   If you are willing to e-mail this info, I will summarize in about
a week for both lists.

   Thank you for your help.
Linda Degenstein
Director UCCRC
Transgenic/ES Cell Facility
The University of Chicago ldeg@midway.uchicago.edu




From YOCKEY at DCSMSERVER.MED.SC.EDU Tue Oct 22 00:15:02 1996
From: YOCKEY at DCSMSERVER.MED.SC.EDU (Courtland E. Yockey)

Subject: selection cassette-dependent phenotypes
Message-ID: <104196F7461@dcsmserver.med.sc.edu>

Hello,

I have a question regarding the phenotypes of targeted mutant mice.

One justification for flanking selection/mutation cassettes with loxP
sites is so that the cassette - and the transcription unit associated
with the selectable marker - can be removed in order to avoid an
artifactual mutant phenotype that is dependent upon its presence.

Does anyone know of published cases in which the phenotype of a mutant
mouse is influenced by the presence of the selectable marker itself
within the mutant locus? Cases in which the phenotype was compared,
for
instance, before and after Cre-mediated excision of a loxP-flanked
cassette?

Thanks for the information.

Courtland Yockey
-----------------------------------
Courtland Yockey, M.S. <yockey@dcsmserver.med.sc.edu>
from the lab of Noriko Shimizu, Ph.D.
                       <shimizu@dcsmserver.med.sc.edu>

University of South Carolina School of Medicine
Department of Developmental Biology & Anatomy
Columbia, SC 29208 USA

Phone# 803-733-1503 <lab>
Fax#   803-733-1533 <dpt>



From Deb_Gumucio.ANATOMY at mailgw.surg.med.umich.edu Tue Oct 22
12:30:59 1996
From: Deb_Gumucio.ANATOMY at mailgw.surg.med.umich.edu (Deb Gumucio)

Subject: selection cassette-depen
Message-ID: <n1366152753.31179@mailgw.surg.med.umich.edu>
        Reply to:   RE>selection cassette-dependent phenotypes

One example is in the beta globin cluster. In 1992, Mark Groudine's lab
showed
that insertion of a neo gene (in the Locus Control Region or LCR)
upstream
from the five clustered globin genes led to severe down-regulation of
the
downstream genes (Genes and Development 6:928). In this case, the
selectable
marker is not in the genes themselves, but in a regulatory region that
is
important for expression of the genes. In more recent work, the same
lab has
inserted neo in place of two of the cores of DNaseI hypersensitive
sites that
comprise the LCR (Genes and Development 9:2203, 1995 and Mol. Cell
Biol.
16:2906, 1996). In each case, the insertion of neo in place of the
core
produced severe effects on globin gene expression, but removing the neo
again
(leaving a core deletion) restored a significant level of globin gene
expression (i.e., it was worse having the neo in than having only the
deletion
itself). The globin cluster may be an unusual case because of the
dependence
of five clustered genes on a remote upstream regulatory unit (the LCR);
the
polar nature of the LCR-promoter interaction appears to be disrupted by
the
presence of the ectopic promoter on neo.

--------------------------------------
Date: 10/21/96 6:18 PM
To: Deb Gumucio
From: transgenic-list@ic.ac.uk
Hello,

I have a question regarding the phenotypes of targeted mutant mice.

One justification for flanking selection/mutation cassettes with loxP
sites is so that the cassette - and the transcription unit associated
with the selectable marker - can be removed in order to avoid an
artifactual mutant phenotype that is dependent upon its presence.

Does anyone know of published cases in which the phenotype of a mutant
mouse is influenced by the presence of the selectable marker itself
within the mutant locus? Cases in which the phenotype was compared,
for
instance, before and after Cre-mediated excision of a loxP-flanked
cassette?

Thanks for the information.

Courtland Yockey
-----------------------------------
Courtland Yockey, M.S. <yockey@dcsmserver.med.sc.edu>
from the lab of Noriko Shimizu, Ph.D.
                       <shimizu@dcsmserver.med.sc.edu>

University of South Carolina School of Medicine
Department of Developmental Biology & Anatomy
Columbia, SC 29208 USA

Phone# 803-733-1503 <lab>
Fax#   803-733-1533 <dpt>



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From:           "Courtland E. Yockey" <YOCKEY@DCSMSERVER.MED.SC.EDU>
Organization: USC School of Medicine
To:             transgenic-list@ic.ac.uk
Date:           Mon, 21 Oct 1996 18:15:02 EST
Subject:        selection cassette-dependent phenotypes
Priority: normal
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From dolle at titus.u-strasbg.fr Tue Oct 22 18:19:24 1996
From: dolle at titus.u-strasbg.fr (Pascal DOLLE)
Subject: selection cassette-dependent phenotypes
Message-ID: <199610221213.OAA08832@titus.u-strasbg.fr >

>Hello,
>
>I have a question regarding the phenotypes of targeted mutant mice.
>
>One justification for flanking selection/mutation cassettes with loxP
>sites is so that the cassette - and the transcription unit associated
>with the selectable marker - can be removed in order to avoid an
>artifactual mutant phenotype that is dependent upon its presence.
>
>Does anyone know of published cases in which the phenotype of a mutant
>mouse is influenced by the presence of the selectable marker itself
>within the mutant locus? Cases in which the phenotype was compared,
for
>instance, before and after Cre-mediated excision of a loxP-flanked
>cassette?
>
>Thanks for the information.
>
>Courtland Yockey
>-----------------------------------

Dear Courtland,

a couple of years ago, we have reported that some 'weird' & dominant
phenotypes in chimeras harboring a Hoxd-10 disruption may have resulted
from integration effect of the neo cassette in the locus. At that time,
the
loxP technique was not developed, so we didn't have this mutation
without
the selection cassette. The ref is Rijli et al., Dev. Dynamics 201, p.
366
(1994).

I guess there will be more and more cases of mutations with or without
neo
cassettes coming out in the next future. You could contact Filippo
Rijli in
our institute who may have other examples of lines with or without TK-
neo
integrations.

Sincerely,



Pascal DOLLE, M.D., Ph.D.

CNRS, Unite 184 INSERM
I.G.B.M.C.
Parc d'innovation
B.P. 163
67404 ILLKIRCH Cedex
FRANCE
Tel: (33) 88 65 33 34 or 40
Fax: (33) 88 65 32 01




From dbowtell at petermac.unimelb.edu.au Tue Oct 22 23:50:38 1996
From: dbowtell at petermac.unimelb.edu.au (David Bowtell)

Subject: selection cassette-dependent phenotypes
In-Reply-To: <199610221213.OAA08832@titus.u-strasbg.fr >
Message-ID: <v03007800ae938c3f52ee@[203.4.164.101]>

As a follow on to the discussion about selectable markers we have
placed a
lacZneo fusion in frame downstream of the Sos1 gene. The marker is
driven
by the Sos1 promoter and there is no heterologous promoter sequences
inserted. We appear to see, however, mosaic expression of lacZ in some
heterozygous animals and wondered about silencing of the targeted locus
in
some tissues. Has anyone else encountered this?

******* Please note new email address **********
David Bowtell
Principal Research Fellow
Research Division
Peter MacCallum Cancer Institute
Locked Bag 1 A'Beckett St,
Melbourne 3000

Lab 61-3-96561287
Office 61-3-96561296
Fax 61-3-96561411
email: dbowtell@petermac.unimelb.edu.au




From anna at screams.gdb.org Wed Oct 23 14:29:45 1996
From: anna at screams.gdb.org (Anna Anagnostopoulos)

Subject: Mycoplasma Infection
Message-ID: <Pine.SOL.3.91.961023092228.11130A-100000@screams.gdb.org>


Hello everyone,

Dr. Yasuzawa is trying to rid his mice of a Mycoplasma infection.   I am
sure he would appreciate additional suggestions.

Thanks a lot

Anna V. Anagnostopoulos
> The reason why I noticed is that my mice are infected   by mycoplasma.
> Especially, the clinical signs of old mice are more     serious than
young.
> I'm worrying about both sudden death and expansion of   contamination.
> I already start to feed tetracyclin to them. But, the   condition has
not
> improved yet.
> If you give me any suggestion how to rescue them, I'm   very happy.
>
>
> yours sincerely,
>
>   Kayoko Yasuzawa
>
>
>
>
>

~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Anna V. Anagnostopoulos, Ph.D.
Division of Biomedical Information Sciences
The Johns Hopkins University School of Medicine
2024 E. Monument Street
Baltimore, MD 21205-2236
tel: (410) 614-3226
fax: (410) 614-0434
e-mail: anna@gdb.org




From milstone at rascal.med.harvard.edu Wed Oct 23 17:26:12 1996
From: milstone at rascal.med.harvard.edu (David Milstone)

Subject: selection cassette-dependent phenotypes
Message-ID: <E0vG5BO-0005ZG-00@juliet.ic.ac.uk>

>Does anyone know of published cases in which the phenotype of a mutant
>mouse is influenced by the presence of the selectable marker itself
>within the mutant locus? Cases in which the phenotype was compared,
for
>instance, before and after Cre-mediated excision of a loxP-flanked
>cassette?
>>
>Courtland Yockey

Selectable markers have been   shown to influence the phenotype of mutant
mice by cis-mediated effects   on transciptional regulation (published
examples include beta globin   LCR element and CD11b). But this is
presumably
distinct from effects of the   coding region sequences of the selectable
marker itself.
David S. Milstone
Vascular Research Division
Department of Pathology
Brigham & Women's Hospital
LMRC 421
221 Longwood Avenue
Boston, MA
USA




From r-carver at nimr.mrc.ac.uk Thu Oct 24 17:40:21 1996
From: r-carver at nimr.mrc.ac.uk (Rick Carver)

Subject: Mycoplasma Infection
Message-ID: <13109.9610231552@nimsn01.nimr.mrc.ac.uk>

Dear All

>Dr. Yasuzawa is trying to rid his mice of a Mycoplasma infection. I
am
>sure he would appreciate additional suggestions.
>
>> The reason why I noticed is that my mice are infected by mycoplasma.
>> Especially, the clinical signs of old mice are more serious than
young.
>> I'm worrying about both sudden death and expansion of contamination.
>> I already start to feed tetracyclin to them. But, the condition has
not
>> improved yet.

My advice would be to rederive them, preferably by embryo transfer.
Antibiosis is a poor option because you can never be certain that it
has
been fully effective and you run a major risk of producing
tetracycline-resistant Mycoplasma. The NRC's 'Infectious Diseases of
Mice
and Rats' has a nice description of the protocol to be followed
post-caesarean rederivation. This protocol is equally valid if you
rederive
by embryo transfer. ET is much more microbiologically secure and if he
is
doing transgenic work he should already have the necessary equipment
and
skills available.

I hope that this is of assistance.




Rick Carver
Head of Biological Services and Institute Veterinarian
National Institute for Medical Research
The Ridgeway
MILL HILL
London                     E-Mail: r-carver@nimr.mrc.ac.uk
NW7 1AA                    Tel: + 44 (0)181 959 3666 ext 2199
United Kingdom             Fax: + 44 (0)181 913 8601




From dknudsen at scruznet.com Thu Oct 24 06:16:18 1996
From: dknudsen at scruznet.com (Dave Knudsen)

Subject: Mycoplasma Infection
Message-ID: <v01540b03ae94a16b55a4@[165.227.113.91]>

>Hello everyone,
>
>Dr. Yasuzawa is trying to rid his mice of a Mycoplasma infection. I
am
>sure he would appreciate additional suggestions.
>
>Thanks a lot
>
>Anna V. Anagnostopoulos
>
>> The reason why I noticed is that my mice are infected by mycoplasma.
>> Especially, the clinical signs of old mice are more serious than
young.
>> I'm worrying about both sudden death and expansion of contamination.
>> I already start to feed tetracyclin to them. But, the condition has
not
>> improved yet.
>> If you give me any suggestion how to rescue them, I'm very happy.

Anna -

I would agree with Rick Carver in that rederivation is the best way to
proceed for a contaminated colony. The assumption under this agreement
is
that the affected colony is a small breeding colony, and not a group of
stock animals that could be eradicated and replaced at the end of an
experiment. I also assume that your colleague, Dr. Yasuzawa, has
completed
the diagnostic process and confirmed mycoplasmosis as the problem. This
diagnosis can be made by serology, culture, or histopathology
(preferably
all three for a colony, to reduce the chance of false negative
findings).
The confirmation itself is important to resolution of the problem - I
can
think of several instances where other opportunistic pathogens,
environmental factors, immunodeficiencies, and/or transgenic phenotypes
have mimicked the clinical presentation of murine mycoplasmosis quite
closely. Also, I would add that antibiosis may help the clinical
appearance
of the animals by temporarily arresting disease progression, but it
will do
nothing in evoking a cure, and treated animals will continue to be
carriers
to infect new introductions (and births) into the colony.

I would personally be most comfortable, however, in dealing with the
rederivation issue by caesarian section; I suspect that confidence is
gained from the method most familiar. Over the years I have had more
positive experiences with caesarian rederivation in eliminating
pathogens
from mouse colonies, and embryo or ovarian manipulations seem always to
break at some point. Perhaps the best method could be determined by
close
questioning of the veterinary staff of the affected institution and
working
with them to resolve the outbreak.

Dave Knudsen DVM, DACLAM
Scotts Valley CA, USA
<dknudsen@scruznet.com)



1996 November
On Mon, 25 Nov 1996, Nick Carboni wrote:

> Hi,
>
> A student mailed me these questions ... maybe one of you could help
him by
> answering these questions or advising him of some references.
>
> >1- About cytoplasmic microinjection using polylysine/DNA mixture:
How
> >polylysine provides cytoplasmic microinjection? Has anyone ever
tried it?
> Is >this technique really revolutionary?
> >
> >2- Do some of you consider it possible to product transgenic mice by
random
> >insertion using ES cells ?
> >
> >3- What about retroviral infection efficiency in the production of
> transgenic & >Ko mice
>
>
> Thank you in advance
>
> Nick
> ------------------------------------------------------------------
> Nick Carboni, VP
> Valoritech
> 1253, McGill College Av., room 620
> Montreal, Quebec, Canada
> H3B 2Y5
> Tel.: (514) 866-8271 / Fax: (514) 866-8272
> E-Mail: carboni@valoritech.com
>
>
>
>




From pinkert at cmed.bhs.uab.edu Tue Nov 26 08:29:43 1996
From: pinkert at cmed.bhs.uab.edu (Dr. C.A. Pinkert)

Subject: Technical questions
Message-ID: <19961126143625396.AAA130@cap1>

> A student mailed me these questions ... maybe one of you could help
him by
> answering these questions or advising him of some references.
>
> >1- About cytoplasmic microinjection using polylysine/DNA mixture:
How
> >polylysine provides cytoplasmic microinjection? Has anyone ever
tried it?
Is >this technique really revolutionary?
It was published - see RL Page et al., Transgenic Research 4:353
(1995).
>
>2- Do some of you consider it possible to product transgenic mice by
random
>insertion using ES cells ?
It is doable - but at issue is whether it is cost-effective compared
to direct DNA microinjection (dependent upon the particular lab ...)?

>3- What about retroviral infection efficiency in the production of
> transgenic & Ko mice
It isn't advantageous - and gene expression issues are not
inconsequential. You can find a goodly number of reviews on the
subject (relative efficiencies, etc.) with a Medline or comparable
abstracting system.

Carl A. Pinkert



From McDonald at WSUHUB.UC.TWSU.EDU Wed Nov 27 14:05:46 1996
From: McDonald at WSUHUB.UC.TWSU.EDU (J. D. McDonald)

Subject: Noise problems and solutions
Message-ID: <01ICC2NFSU7M0A3LB8@WSUHUB.UC.TWSU.EDU>

I am the director of our university's animal care facility and I would
like
to call upon the mouse husbandry expertise of this group with regard to
a
specific noise issue. Our building will soon be undergoing an
extensive
upgrading of the HVAC (heating, ventilation, and air conditioning)
system.
This will not occur within the cluster of rooms that comprise our
animal
care facility but will be occurring all around it for a period of about
a
month. We are anticipating significant amounts of intermittent
construction
noise and are investigating ways of dealing with this so as to minimize
the
stress levels on our research animals (primarily mice). In discussing
options, one idea is to introduce some type of white noise to mask the
construction noise. Does anyone out there have any experience with
this
strategy? Can you point me to pertinent literature (a quick Medline
search
didn't turn up anything of apparent value)? Do you know of other more
effective ways of dealing with this problem? Please send any advice
that
will likely be helpful by email. If I get a good number of responses,
I
will post a summary.

J. David McDonald, Ph. D., Assistant Professor
Wichita State University
Department of Biological Sciences
1845 Fairmount, Box #26
Wichita, KS 67260-0026
Telephone 316-978-3111 x16
Fax 316-978-3772
Voicemail 316-978-3978 x6097
Email mcdonald@wsuhub.uc.twsu.edu




From crussell at ResGen.COM Wed Nov 27 16:11:47 1996
From: crussell at ResGen.COM (Chris Russell)

Subject: Technical questions
Message-ID: <199611271611.KAA27982@twister.resgen.com>

Further to Dr. Pinkert's answers to the following question:

>>1- About cytoplasmic microinjection using polylysine/DNA mixture: How
>>polylysine provides cytoplasmic microinjection? Has anyone ever tried
it?
>Is >this technique really revolutionary?

R. L. Page, S. P. Butler, A. Subramanian, F. C. Gwazdauskas, J. L.
Johnson,
and W. H. Velander. 1995. "Transgenesis in Mice by Cytoplasmic
Injection of
Polylysine/DNA Mixtures". Transgenic Research 4:353-360


The polyLysine-DNA (pLDNA) mixture is injected into the cytoplasm of
fertilized embryos, similar to the technique of pronuclear
microinjection.
However, in this case, it is not necessary to hit nor even see the
pronucleus. The technique is not a efficient as pronuclear
microinjection
-- about 15% maximum transgenesis observed in mice -- but does give
rise to
functional, expressing transgenic animals, which seemed to be
equivalent to
transgenic mice made by pronuclear microinjection. It also provides
another
technique in the arsenal of transgenic animal production.

Is the technique revolutionary? Depends on your opinion of
demonstration of
new techniques. Apparently the US patent office thought so.    The
patent
was awarded to Page, Velander, and perhaps one other author. To the
best of
my knowledge, PPL, Inc (of Blacksburg, VA and Edinburgh, Scotland)
holds the
exclusive license to cytoplasmic injection for generation of transgenic
animals.

Let me know if there are further questions, and I'd be glad to try to
help
(I was in the transgenic research group at Virginia Tech at the time of
development of the technique).

Chris
***********************************************************************
****
                                 Christopher G. Russell, Ph.D.
                                 RESEARCH GENETICS, INC
                                     Resources for Research
-_+            _+_+           ++-_         _+-_         _+_          _++
_+-_
     -+        -+     -+       +-    +-      -+   +       -+   -+         -+    +
-+
      -+     -+       -+      +-     -+     -+    -+     -+     -+       -+
-+      -+
         -++-+         -+_-+         -+_+        -++-          -+_++         -+_+
                             2700 Memorial Parkway SW
                                    Huntsville, AL 35801
 Phone 1-800-533-4363, ext 2211 or Direct 1-800-711-2089
                                     U.K. 0-800-89-1393
                  Fax 1-205-551-1021 or 1-205-536-9016
                                    crussell@resgen.com
***********************************************************************
******




From tonyjames at cuhk.edu.hk Fri Nov 29 02:40:51 1996
From: tonyjames at cuhk.edu.hk (A E James)
Subject: Noise problems and solutions
Message-ID: <199611290240.KAA25003@hpg50a.csc.cuhk.edu.hk>


Can I suggest you correspond with Dr. Gerald Glough in the UK, he is a
recognised expert on these matters.
A relevant paper is Enviromental Effects on Animals used in Biomedical
Research.
G. Clough, Biol. Rev. Vol. 57,pp. 487-523
Gerald can be contacted through Alanan Consultancy Sevives PO Box 230
York
YO1 1GG
England, ph/fx 44-1904-656368


>I am the director of our university's animal care facility and I would
like
>to call upon the mouse husbandry expertise of this group with regard
to a
>specific noise issue. Our building will soon be undergoing an
extensive
>upgrading of the HVAC (heating, ventilation, and air conditioning)
system.
>This will not occur within the cluster of rooms that comprise our
animal
>care facility but will be occurring all around it for a period of
about a
>month. We are anticipating significant amounts of intermittent
construction
>noise and are investigating ways of dealing with this so as to
minimize the
>stress levels on our research animals (primarily mice). In discussing
>options, one idea is to introduce some type of white noise to mask the
>construction noise. Does anyone out there have any experience with
this
>strategy? Can you point me to pertinent literature (a quick Medline
search
>didn't turn up anything of apparent value)? Do you know of other more
>effective ways of dealing with this problem? Please send any advice
that
>will likely be helpful by email. If I get a good number of responses,
I
>will post a summary.
>
>J. David McDonald, Ph. D., Assistant Professor
>Wichita State University
>Department of Biological Sciences
>1845 Fairmount, Box #26
>Wichita, KS 67260-0026
>Telephone 316-978-3111 x16
>Fax 316-978-3772
>Voicemail 316-978-3978 x6097
>Email mcdonald@wsuhub.uc.twsu.edu
>
>
>
>
Tony James
BVSc MACVSc
Director
Laboratory Animal Unit
Chinese University of Hong Kong
Shatin, New Territories, Hong Kong
ph: 852 2609 6862
fx: 852 2603 5723
email: tonyjames@cuhk.edu.hk
.....................................
"nothing great was achieved without enthusiasm."
R. W. Emerson




1996 December
From pasceri at sickkids.on.ca Mon Dec 2 17:05:25 1996
From: pasceri at sickkids.on.ca (Peter Pasceri)
Date: Wed Aug 20 11:52:56 2003
Subject: Seasonal changes
Message-ID: <v01510101aec8f1d409f8@[142.20.36.217]>


I have a question which is not pressing but would appreciate any input
from
those of you that have experienced the same.

Over the past three years I have noticed that my animals seem to behave
differently during the months of November and December. They give less
eggs when superovulated and there seems to be more resorbtions than is
acceptable. Last week more than 80% resorbtions !

Culture media supports growth in vitro and is not suspected.    Things
usually straighten out by late January.

Any ideas?

Peter


--
 Peter Pasceri
 pasceri@sickkids.on.ca
 Genetics Research
 The Hospital for Sick Children
 Toronto, Ontario,Canada
 Phone: ( 416 ) 813-8169




From gotto at leland.Stanford.EDU   Mon Dec   2 16:44:22 1996
From: gotto at leland.Stanford.EDU (Glen Otto)
Date: Wed Aug 20 11:52:56 2003
Subject: Seasonal changes
In-Reply-To: <v01510101aec8f1d409f8@[142.20.36.217]>
Message-ID: <v03007800aec90feaa24a@[171.65.18.36]>

>Over the past three years I have noticed that my animals seem to
behave
>differently during the months of November and December. They give
less
>eggs when superovulated and there seems to be more resorbtions than is
>acceptable. Last week more than 80% resorbtions !


Statistically significant annual cycles in mouse reproductive
efficiency
(even in climate-controlled indoor facilities) is not uncommon in
populations large enough to notice it, and I've been told that some
major
breeding facilities take it into account when making production
projections.

I'm not sure anybody knows why, and when I mentioned it to a prominent
sociobiologist a few years ago she thought it would make a great thesis
study. Maybe a fallback project for someone who can't get a strain made
because the mice don't produce!

glen

_________________________________________
Glen Otto, DVM
Dept. of Comparative Medicine               gotto@leland.stanford.edu
Stanford University
Quad 7, Bldg. 330                           415/723-3876   voice
Stanford, CA 94305-5410                     415/725-0940   fax




From jklohse at facstaff.wisc.edu Mon Dec 2 22:08:33 1996
From: jklohse at facstaff.wisc.edu (Jan K. Lohse)
Date: Wed Aug 20 11:52:56 2003
Subject: Seasonal changes
In-Reply-To: <v01510101aec8f1d409f8@[142.20.36.217]>
Message-ID: <v03007800aec9641a3bf2@[144.92.48.83]>

Many workers with rodent embryos experience some seasonal variation.
This
is sometimes a slump in early winter, as you described, and sometimes
one
problem period around October & another around April, when the weather
is
changing the most. This occurs even in light/temperature/humidity
controlled colonies. I have never heard of a good, conclusive study
tracking down the reasons for this, but it seems to be fairly common.
From browng at medicine.wustl.edu Tue Dec 3 07:09:24 1996
From: browng at medicine.wustl.edu (Gary Brown)
Date: Wed Aug 20 11:52:56 2003
Subject: Seasonal changes
In-Reply-To: <v01510101aec8f1d409f8@[142.20.36.217]>
Message-ID: <Pine.GSO.3.94.961203065320.1467A-100000@medicine>

Hi!

Just my $0.02 worth... I too have experienced such seasonal phenomena
in
the past, and although might give a probable cause I can't pretend the
answer will be easy to implement! In general, the most difficult times
of
year coincide with the late phase of exterior climactic change ie Fall
and
Spring. The building in which animals are housed will undergo its own
climactic adjustments correspondingly as air conditioning, humidity
control and heating readjust. The efficiency (often times the age) of
the
building in making this transition will determine the severity of the
observed effect as a new environmental stress is added to post surgical
mice at this time. All I can suggest is that you work with your
building
manager / facility manager to minimize the impact of such changes, and
maintain accurate records of high/low humidity, temp etc. to visualise
if
this is indeed the problem.

Another small(?) consideration. In my own situation, we were
originally
housed in the basement area of the local Childrens Hospital until very
recently - we've since moved to a purpose built facility :-) In this
situation, the environmental controls were shared between our mouse
floor
and the one above (a "people" floor), necessitating a less-than-
desirable
compromise. Nothing's ever easy......

Regards,

Gary Brown
Email: browng@medicine.wustl.edu or 75017.1050@compuserve.com
Web: http://ourworld.compuserve.com/homepages/TheBroons/
         "The Microinjection Workshop"


On Mon, 2 Dec 1996, Peter Pasceri wrote:

>
> I have a question which is not pressing but would appreciate any
input from
> those of you that have experienced the same.
>
> Over the past three years I have noticed that my animals seem to
behave
> differently during the months of November and December. They give
less
> eggs when superovulated and there seems to be more resorbtions than
is
> acceptable. Last week more than 80% resorbtions !
>
> Culture media supports growth in vitro and is not suspected. Things
> usually straighten out by late January.
>
> Any ideas?
>
> Peter
>
>
> --
> Peter Pasceri
> pasceri@sickkids.on.ca
> Genetics Research
> The Hospital for Sick Children
> Toronto, Ontario,Canada
> Phone: ( 416 ) 813-8169
>
>
>
>
>




From tjf at uci.edu Tue Dec 3 11:43:34 1996
From: tjf at uci.edu (Tom Fielder)
Date: Wed Aug 20 11:52:56 2003
Subject: in vitro vs. in vivo expression
Message-ID: <v03007807aec9c355cfed@[128.200.21.145]>

I would like to get some feedback from those of you involved in
transgenic
research (this will be posted on Embryo Mail and Compmed as well). I
am
running a new transgenic core facility at UC-Irvine. We are debating
whether or not to require some evidence of protein expression from DNA
constructs before doing pronuclear injection. On the one hand, it
would be
nice to know that the construct has at least some chance of working and
that the investigator didn't make a mistake in the engineering. On the
other hand, some people argue that in vitro expression cannot guarantee
in
vivo expression (certainly true) and is in fact not even a decent
predictor
of in vivo success. What do people require in terms of construct
testing
(e.g., RE digest, gel picture, sequencing, in vitro expression), and
does
anyone have any statistics on the predictability of in vivo expression
from
in vitro results? If I get enough responses, I'll summarize for the
list.
Thanks for your help!
Tom

Thomas J. Fielder
UC-Irvine
University Lab Animal Resources
147 BSA
Irvine, CA 92697-1310
e-mail: tjf@uci.edu
phone: 714-824-8579
FAX: 714-824-2003




From david.threadgill at mcmail.vanderbilt.edu Thu Dec 5 10:00:49
1996
From: david.threadgill at mcmail.vanderbilt.edu
(david.threadgill@mcmail.vanderbilt.edu)
Date: Wed Aug 20 11:52:56 2003
Subject: spretus
Message-ID: <9611058498.AA849808822@in2.mcmail.vanderbilt.edu>

Does anyone have experience with superovulating spretus females? Do
they respond
to the same level of hormones as musculus? Do spretus males mate
reliably with
superovulated females?
Thanks,
David Threadgill
Dept of Cell Biology
Vanderbilt Univ
Nashville, TN
david.threadgill@mcmail.vanderbilt.edu




From p.sobieszczuk at ic.ac.uk Fri Dec 6 17:40:36 1996
From: p.sobieszczuk at ic.ac.uk (Peter Sobieszczuk)
Date: Wed Aug 20 11:52:56 2003
Subject: Seasonal changes/Forwarded from Pete Willan
<pcw3@leicester.ac.uk>
Message-ID: <v01510116aece0b57a729@[155.198.45.54]>

>From: Pete Willan <pcw3@leicester.ac.uk>
>To: transgenic-list@ic.ac.uk
>Date: Thu, 5 Dec 1996 09:49:20 +0100 (BST)
>Subject: Re: Seasonal changes
>Reply-to: "Division of Biomedical Services"@leicester.ac.uk,
>          University@leicester.ac.uk
>CC: DF9@leicester.ac.uk
>Priority: normal
>X-mailer: Pegasus Mail for Windows (v2.40)
>Message-ID: <32686861B3@lupin.le.ac.uk>
>
>Hi to all
>
>This may not be the most authentic answer to reduction in breeding
>colonies and embryo reabsorbtion(I am assuming that changes in time
clocks
>have also been investigated) has anyone out there
>looked at the following senario...
>
>
>The only variation into the breeding colony that may effect the
>animals could be born(exuse pun) from the effect of seasonal
>changes on the Animal Welfare Staff being transmitted somehow
>to the animals.
>
>I would be interested in comments.
>
>Seasonal Greetings to all!
>
>Pete Willan




From p.sobieszczuk at ic.ac.uk Sat Dec 7 16:22:37 1996
From: p.sobieszczuk at ic.ac.uk (Peter Sobieszczuk)
Date: Wed Aug 20 11:52:56 2003
Subject: Noise problems/Forwarded from
rap@vesalius.Anatomy.Unimelb.EDU.AU
Message-ID: <v01510113aecf4901edb5@[155.198.45.54]>

>To: transgenic-list@ic.ac.uk
>From: rap@vesalius.Anatomy.Unimelb.EDU.AU (Rachael Parkinson)
>Subject: Re: Noise problems
>Content-Length: 718
>
>My department is currently undergoing construction work and, yes, this
has
>had quite an adverse effect on mouse breeding and cannibalism of mouse
>pups. I have no good news. We had our "temporary" mouse room
>sound-proofed to try to minimalise the noise levels in there, but the
>construction work has caused quite serious vibrations throughout the
>building which I believe to be the primary cause of my animals'
stress. If
>your building works are expected to cause vibrations, then I think
this
>will be more of a concern than noise which may be able to be masked
(by
>white noise as you have suggested).
>
>Rachael Parkinson
>Department Of Anatomy and Cell Biology
>University of Melbourne
>Parkville VIC 3052
>Australia.




From BIOGEN at aol.com Sat Dec 7 13:51:07 1996
From: BIOGEN at aol.com (BIOGEN@aol.com)
Date: Wed Aug 20 11:52:56 2003
Subject: Noise problems/Forwarded from
rap@vesalius.Anatomy.Unimelb.EDU.AU
Message-ID: <961207135106_1153114923@emout13.mail.aol.com>

My first interaction with the group is somewhat off-the-wall: In
regard to
Rachel's observation of the decline in breeding and the associated
cannabalism as a result of the auditory assault generated from
construction
work-- Would you please comment on the estimated percentile of decline
in
breeding and/or the possible general effect this may have on the
population
in general? And, do you have any way of quantifying the decibel level
and
mean frequency of the noise?

If these seem like strange requests, upon further consideration you
might see
where I'm going: If the decline in breeding is statistically large,
and the
frequencies involved can be isolated into a range minimally obtrusive
to
humans, it might be of use in eradication or in the prevention of
infestation
in structures. As my primary education was in electronic engineering,
I am
always curious about the possibilities of interaction between
disciplines
(biology and electronics). Thanks for your consideration!

Kind Regards,

Lou Jones / Biogenics



From tjf at uci.edu Wed Dec 11 15:25:19 1996
From: tjf at uci.edu (Tom Fielder)
Date: Wed Aug 20 11:52:57 2003
Subject: in vitro expression testing
Message-ID: <v03007805aed48328c92d@[128.200.21.145]>

Here is a summary of the replies I received in response to my question
regarding the advisibility of in vitro expression testing of DNA
constructs
prior to pronuclear injection:
There were 10 respondents. Apparently no one demands mandatory in
vitro
testing, although 3 respondents said they ask for or encourage such
testing
when possible (e.g., if an appropriate cell line is available). Of the
seven replies which listed what they do require, the following items
were
mentioned:

gel photo: 6
restriction digest: 2
sequence data around cloning sites:   2
evidence of specific PCR assay: 1

Thanks very much for your replies.

Tom

Thomas J. Fielder
UC-Irvine
University Lab Animal Resources
147 BSA
Irvine, CA 92697-1310
e-mail: tjf@uci.edu
phone: 714-824-8579
FAX: 714-824-2003




From i.rosewell at icrf.icnet.uk Tue Dec 17 13:01:45 1996
From: i.rosewell at icrf.icnet.uk (Ian -Transgenic Unit)
Date: Wed Aug 20 11:52:57 2003
Subject: transmission?
Message-ID: <v01530506aedc393371c8@[143.65.49.6]>

Hello

We have recently achieved germline transmission from 2 independent
clones.
The clones were from seperate knockout projects and seperate ES cell
lines.
The clones transmited poorly. In the first case after a number of
litters one chimera gave rise to what was initially identified to me as
a
chimera, the coat colour which should have been( Bl6 x 129) agouti was
uneven with areas of black fur. The chimera that gave rise to this
then
went on to father a true agouti and het. mouse. The first mouse was
negative. This has happened again. Can anyone provide an explanation?

Thanks
From anna at screams.gdb.org Thu Dec 19 09:25:39 1996
From: anna at screams.gdb.org (Anna Anagnostopoulos)
Date: Wed Aug 20 11:52:57 2003
Subject: purchase/breeding of APO E KO mice
In-Reply-To: <aeb728ea03021004b24e@[130.225.37.173]>
Message-ID: <Pine.SOL.3.91.961219090944.11285B-100000@screams.gdb.org>

On Tue, 19 Nov 1996, Frederik =?iso-8859-1?Q?Dagn=E6s=2DHansen?= wrote:

Dear Frederik,

I don't know much about availability in Europe. However, Apoe strains
C57BL/6J-Apoe(tm1Unc) generated by Dr. Nobuyo Maeda and strain B6,
129-Apoe(tm1Unc) Ldlr(tm1Her) generated by Dr. Joachim Herz are both
available at the Induced Mutant Resources at the Jackson
Laboratories, Bar Harbor, Maine, USA (Stock #s are JR2052 and JR2245,
respectively). The latter is of limited
distribution. You may check the IMR Web site for additional
information
(http://lena.jax.org/resources/documents/imr/). I can also provide the
following contact information to you through TBASE
(http://www.gdb.org/Dan/tbase/tbase.html):

Dr. Nobuyo Maeda

Univ. of North Carolina at Chaperl HIll
Dept of Pathology
CB7525
Brinkhous-Bullitt Building
Chaperl Hill, NC 29599-7525
(608) 262-1047


You may also want to contact:

Dr. Jan Breslow
The Rockefeller University
Biochemical Genetics and Metabolism
Box 29 HOS
1230 York Ave,
New York, NY 10021
(212) 570-7700

They may answer some of your questions regarding availability and
breeding.

I hope this will be of some assistance

Sincerely

Anna V. Anagnostopoulos
e-mail: anna@gdb.org

 > Hej everybody >
> A researcher working in cardiovascular research whishes to work with
the
> APO E knockout mice (C57BL/6J-APO E KO ) .
> As it is difficult to purchase these mice from commercial breeders in
> Europe, he has tryed to establish a small breeding colony of these.
> The outcome is very bad as the mothers eat the pups just after
delivery.
> Does anyone have some good ideas how to get these mice, how to make
them
> breed???????+
>
> best regards
>
>   Frederik Dagn�s-Hansen
>
>   Frederik Dagnaes-Hansen,D.V.M., Ph.D.
>   Inst. Med.Microbiol.& Imm.
>   Bartholin Building, Building 240,
>   University of Aarhus,
>   8000 Aarhus C,
>   Tel + 45 89 42 1774
>   Fax + 45 8819 6128
>   Email: mikrfdh@svfcd.aau.dk
>
>
>
>
>
>

~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Anna V. Anagnostopoulos, Ph.D.
Division of Biomedical Information Sciences
The Johns Hopkins University School of Medicine
2024 E. Monument Street
Baltimore, MD 21205-2236
tel: (410) 614-3226
fax: (410) 614-0434
e-mail: anna@gdb.org




From tjf at uci.edu Mon Dec 30 10:49:02 1996
From: tjf at uci.edu (Tom Fielder)
Date: Wed Aug 20 11:52:57 2003
Subject: Balb/C knockouts
Message-ID: <v03007802aeed4f41ca8f@[128.200.21.145]>

Does anyone know of a source of CD4 and/or CD8 knockouts in Balb/C
mice? A
colleague of mine has contacted Taconic and Jax and they only have them
in
C57's. Thanks in advance.
Tom
Thomas J. Fielder
UC-Irvine
University Lab Animal Resources
147 BSA
Irvine, CA 92697-1310
e-mail: tjf@uci.edu
phone: 714-824-8579
FAX: 714-824-2003




From pasceri at sickkids.on.ca Mon Dec 2 21:05:25 1996
From: pasceri at sickkids.on.ca (Peter Pasceri)

Subject: Seasonal changes
Message-ID: <v01510101aec8f1d409f8@[142.20.36.217]>


I have a question which is not pressing but would appreciate any input
from
those of you that have experienced the same.

Over the past three years I have noticed that my animals seem to behave
differently during the months of November and December. They give less
eggs when superovulated and there seems to be more resorbtions than is
acceptable. Last week more than 80% resorbtions !

Culture media supports growth in vitro and is not suspected.   Things
usually straighten out by late January.

Any ideas?

Peter


--
 Peter Pasceri
 pasceri@sickkids.on.ca
 Genetics Research
 The Hospital for Sick Children
 Toronto, Ontario,Canada
 Phone: ( 416 ) 813-8169




From gotto at leland.Stanford.EDU Tue Dec 3 00:44:22 1996
From: gotto at leland.Stanford.EDU (Glen Otto)

Subject: Seasonal changes
In-Reply-To: <v01510101aec8f1d409f8@[142.20.36.217]>
Message-ID: <v03007800aec90feaa24a@[171.65.18.36]>
>Over the past three years I have noticed that my animals seem to
behave
>differently during the months of November and December. They give
less
>eggs when superovulated and there seems to be more resorbtions than is
>acceptable. Last week more than 80% resorbtions !


Statistically significant annual cycles in mouse reproductive
efficiency
(even in climate-controlled indoor facilities) is not uncommon in
populations large enough to notice it, and I've been told that some
major
breeding facilities take it into account when making production
projections.

I'm not sure anybody knows why, and when I mentioned it to a prominent
sociobiologist a few years ago she thought it would make a great thesis
study. Maybe a fallback project for someone who can't get a strain made
because the mice don't produce!

glen

_________________________________________
Glen Otto, DVM
Dept. of Comparative Medicine               gotto@leland.stanford.edu
Stanford University
Quad 7, Bldg. 330                           415/723-3876   voice
Stanford, CA 94305-5410                     415/725-0940   fax




From jklohse at facstaff.wisc.edu Tue Dec 3 05:08:33 1996
From: jklohse at facstaff.wisc.edu (Jan K. Lohse)

Subject: Seasonal changes
In-Reply-To: <v01510101aec8f1d409f8@[142.20.36.217]>
Message-ID: <v03007800aec9641a3bf2@[144.92.48.83]>

Many workers with rodent embryos experience some seasonal variation.
This
is sometimes a slump in early winter, as you described, and sometimes
one
problem period around October & another around April, when the weather
is
changing the most. This occurs even in light/temperature/humidity
controlled colonies. I have never heard of a good, conclusive study
tracking down the reasons for this, but it seems to be fairly common.




From browng at medicine.wustl.edu Tue Dec 3 13:09:24 1996
From: browng at medicine.wustl.edu (Gary Brown)
Subject: Seasonal changes
In-Reply-To: <v01510101aec8f1d409f8@[142.20.36.217]>
Message-ID: <Pine.GSO.3.94.961203065320.1467A-100000@medicine>

Hi!

Just my $0.02 worth... I too have experienced such seasonal phenomena
in
the past, and although might give a probable cause I can't pretend the
answer will be easy to implement! In general, the most difficult times
of
year coincide with the late phase of exterior climactic change ie Fall
and
Spring. The building in which animals are housed will undergo its own
climactic adjustments correspondingly as air conditioning, humidity
control and heating readjust. The efficiency (often times the age) of
the
building in making this transition will determine the severity of the
observed effect as a new environmental stress is added to post surgical
mice at this time. All I can suggest is that you work with your
building
manager / facility manager to minimize the impact of such changes, and
maintain accurate records of high/low humidity, temp etc. to visualise
if
this is indeed the problem.

Another small(?) consideration. In my own situation, we were
originally
housed in the basement area of the local Childrens Hospital until very
recently - we've since moved to a purpose built facility :-) In this
situation, the environmental controls were shared between our mouse
floor
and the one above (a "people" floor), necessitating a less-than-
desirable
compromise. Nothing's ever easy......

Regards,

Gary Brown
Email: browng@medicine.wustl.edu or 75017.1050@compuserve.com
Web: http://ourworld.compuserve.com/homepages/TheBroons/
         "The Microinjection Workshop"


On Mon, 2 Dec 1996, Peter Pasceri wrote:

>
> I have a question which is not pressing but would appreciate any
input from
> those of you that have experienced the same.
>
> Over the past three years I have noticed that my animals seem to
behave
> differently during the months of November and December. They give
less
> eggs when superovulated and there seems to be more resorbtions than
is
> acceptable. Last week more than 80% resorbtions !
>
> Culture media supports growth in vitro and is not suspected. Things
> usually straighten out by late January.
>
> Any ideas?
>
> Peter
>
>
> --
> Peter Pasceri
> pasceri@sickkids.on.ca
> Genetics Research
> The Hospital for Sick Children
> Toronto, Ontario,Canada
> Phone: ( 416 ) 813-8169
>
>
>
>
>




From tjf at uci.edu Tue Dec 3 19:43:34 1996
From: tjf at uci.edu (Tom Fielder)

Subject: in vitro vs. in vivo expression
Message-ID: <v03007807aec9c355cfed@[128.200.21.145]>

I would like to get some feedback from those of you involved in
transgenic
research (this will be posted on Embryo Mail and Compmed as well). I
am
running a new transgenic core facility at UC-Irvine. We are debating
whether or not to require some evidence of protein expression from DNA
constructs before doing pronuclear injection. On the one hand, it
would be
nice to know that the construct has at least some chance of working and
that the investigator didn't make a mistake in the engineering. On the
other hand, some people argue that in vitro expression cannot guarantee
in
vivo expression (certainly true) and is in fact not even a decent
predictor
of in vivo success. What do people require in terms of construct
testing
(e.g., RE digest, gel picture, sequencing, in vitro expression), and
does
anyone have any statistics on the predictability of in vivo expression
from
in vitro results? If I get enough responses, I'll summarize for the
list.
Thanks for your help!
Tom

Thomas J. Fielder
UC-Irvine
University Lab Animal Resources
147 BSA
Irvine, CA 92697-1310
e-mail: tjf@uci.edu
phone: 714-824-8579
FAX: 714-824-2003




From david.threadgill at mcmail.vanderbilt.edu Thu Dec   5 16:00:49
1996
From: david.threadgill at mcmail.vanderbilt.edu
(david.threadgill@mcmail.vanderbilt.edu)

Subject: spretus
Message-ID: <9611058498.AA849808822@in2.mcmail.vanderbilt.edu>

Does anyone have experience with superovulating spretus females? Do
they respond
to the same level of hormones as musculus? Do spretus males mate
reliably with
superovulated females?
Thanks,
David Threadgill
Dept of Cell Biology
Vanderbilt Univ
Nashville, TN
david.threadgill@mcmail.vanderbilt.edu




From p.sobieszczuk at ic.ac.uk Fri Dec 6 17:40:36 1996
From: p.sobieszczuk at ic.ac.uk (Peter Sobieszczuk)

Subject: Seasonal changes/Forwarded from Pete Willan
<pcw3@leicester.ac.uk>
Message-ID: <v01510116aece0b57a729@[155.198.45.54]>

>From: Pete Willan <pcw3@leicester.ac.uk>
>To: transgenic-list@ic.ac.uk
>Date: Thu, 5 Dec 1996 09:49:20 +0100 (BST)
>Subject: Re: Seasonal changes
>Reply-to: "Division of Biomedical Services"@leicester.ac.uk,
>          University@leicester.ac.uk
>CC: DF9@leicester.ac.uk
>Priority: normal
>X-mailer: Pegasus Mail for Windows (v2.40)
>Message-ID: <32686861B3@lupin.le.ac.uk>
>
>Hi to all
>
>This may not be the most authentic answer to reduction in breeding
>colonies and embryo reabsorbtion(I am assuming that changes in time
clocks
>have also been investigated) has anyone out there
>looked at the following senario...
>
>
>The only variation into the breeding colony that may effect the
>animals could be born(exuse pun) from the effect of seasonal
>changes on the Animal Welfare Staff being transmitted somehow
>to the animals.
>
>I would be interested in comments.
>
>Seasonal Greetings to all!
>
>Pete Willan




From p.sobieszczuk at ic.ac.uk Sat Dec 7 16:22:37 1996
From: p.sobieszczuk at ic.ac.uk (Peter Sobieszczuk)

Subject: Noise problems/Forwarded from
rap@vesalius.Anatomy.Unimelb.EDU.AU
Message-ID: <v01510113aecf4901edb5@[155.198.45.54]>

>To: transgenic-list@ic.ac.uk
>From: rap@vesalius.Anatomy.Unimelb.EDU.AU (Rachael Parkinson)
>Subject: Re: Noise problems
>Content-Length: 718
>
>My department is currently undergoing construction work and, yes, this
has
>had quite an adverse effect on mouse breeding and cannibalism of mouse
>pups. I have no good news. We had our "temporary" mouse room
>sound-proofed to try to minimalise the noise levels in there, but the
>construction work has caused quite serious vibrations throughout the
>building which I believe to be the primary cause of my animals'
stress. If
>your building works are expected to cause vibrations, then I think
this
>will be more of a concern than noise which may be able to be masked
(by
>white noise as you have suggested).
>
>Rachael Parkinson
>Department Of Anatomy and Cell Biology
>University of Melbourne
>Parkville VIC 3052
>Australia.
From BIOGEN at aol.com Sat Dec 7 18:51:07 1996
From: BIOGEN at aol.com (BIOGEN@aol.com)
Date: Fri Jul 9 14:03:00 2004
Subject: Noise problems/Forwarded from
rap@vesalius.Anatomy.Unimelb.EDU.AU
Message-ID: <961207135106_1153114923@emout13.mail.aol.com>

My first interaction with the group is somewhat off-the-wall: In
regard to
Rachel's observation of the decline in breeding and the associated
cannabalism as a result of the auditory assault generated from
construction
work-- Would you please comment on the estimated percentile of decline
in
breeding and/or the possible general effect this may have on the
population
in general? And, do you have any way of quantifying the decibel level
and
mean frequency of the noise?

If these seem like strange requests, upon further consideration you
might see
where I'm going: If the decline in breeding is statistically large,
and the
frequencies involved can be isolated into a range minimally obtrusive
to
humans, it might be of use in eradication or in the prevention of
infestation
in structures. As my primary education was in electronic engineering,
I am
always curious about the possibilities of interaction between
disciplines
(biology and electronics). Thanks for your consideration!

Kind Regards,

Lou Jones / Biogenics



From tjf at uci.edu Wed Dec 11 23:25:19 1996
From: tjf at uci.edu (Tom Fielder)
Date: Fri Jul 9 14:03:00 2004
Subject: in vitro expression testing
Message-ID: <v03007805aed48328c92d@[128.200.21.145]>

Here is a summary of the replies I received in response to my question
regarding the advisibility of in vitro expression testing of DNA
constructs
prior to pronuclear injection:

There were 10 respondents. Apparently no one demands mandatory in
vitro
testing, although 3 respondents said they ask for or encourage such
testing
when possible (e.g., if an appropriate cell line is available). Of the
seven replies which listed what they do require, the following items
were
mentioned:

gel photo: 6
restriction digest: 2
sequence data around cloning sites:   2
evidence of specific PCR assay: 1

Thanks very much for your replies.

Tom

Thomas J. Fielder
UC-Irvine
University Lab Animal Resources
147 BSA
Irvine, CA 92697-1310
e-mail: tjf@uci.edu
phone: 714-824-8579
FAX: 714-824-2003




From i.rosewell at icrf.icnet.uk Tue Dec 17 12:01:45 1996
From: i.rosewell at icrf.icnet.uk (Ian -Transgenic Unit)
Date: Fri Jul 9 14:03:00 2004
Subject: transmission?
Message-ID: <v01530506aedc393371c8@[143.65.49.6]>

Hello

We have recently achieved germline transmission from 2 independent
clones.
The clones were from seperate knockout projects and seperate ES cell
lines.
The clones transmited poorly. In the first case after a number of
litters one chimera gave rise to what was initially identified to me as
a
chimera, the coat colour which should have been( Bl6 x 129) agouti was
uneven with areas of black fur. The chimera that gave rise to this
then
went on to father a true agouti and het. mouse. The first mouse was
negative. This has happened again. Can anyone provide an explanation?

Thanks




From anna at screams.gdb.org Thu Dec 19 14:25:39 1996
From: anna at screams.gdb.org (Anna Anagnostopoulos)
Date: Fri Jul 9 14:03:00 2004
Subject: purchase/breeding of APO E KO mice
In-Reply-To: <aeb728ea03021004b24e@[130.225.37.173]>
Message-ID: <Pine.SOL.3.91.961219090944.11285B-100000@screams.gdb.org>

On Tue, 19 Nov 1996, Frederik =?iso-8859-1?Q?Dagn=E6s=2DHansen?= wrote:

Dear Frederik,

I don't know much about availability in Europe. However, Apoe strains
C57BL/6J-Apoe(tm1Unc) generated by Dr. Nobuyo Maeda and strain B6,
129-Apoe(tm1Unc) Ldlr(tm1Her) generated by Dr. Joachim Herz are both
available at the Induced Mutant Resources at the Jackson
Laboratories, Bar Harbor, Maine, USA (Stock #s are JR2052 and JR2245,
respectively). The latter is of limited
distribution. You may check the IMR Web site for additional
information
(http://lena.jax.org/resources/documents/imr/). I can also provide the
following contact information to you through TBASE
(http://www.gdb.org/Dan/tbase/tbase.html):

Dr. Nobuyo Maeda

Univ. of North Carolina at Chaperl HIll
Dept of Pathology
CB7525
Brinkhous-Bullitt Building
Chaperl Hill, NC 29599-7525
(608) 262-1047


You may also want to contact:

Dr. Jan Breslow
The Rockefeller University
Biochemical Genetics and Metabolism
Box 29 HOS
1230 York Ave,
New York, NY 10021
(212) 570-7700

They may answer some of your questions regarding availability and
breeding.

I hope this will be of some assistance

Sincerely

Anna V. Anagnostopoulos
e-mail: anna@gdb.org

 > Hej everybody >
> A researcher working in cardiovascular research whishes to work with
the
> APO E knockout mice (C57BL/6J-APO E KO ) .
> As it is difficult to purchase these mice from commercial breeders in
> Europe, he has tryed to establish a small breeding colony of these.
> The outcome is very bad as the mothers eat the pups just after
delivery.
> Does anyone have some good ideas how to get these mice, how to make
them
> breed???????+
>
> best regards
>
>   Frederik Dagn�s-Hansen
>
>   Frederik Dagnaes-Hansen,D.V.M., Ph.D.
>   Inst. Med.Microbiol.& Imm.
>   Bartholin Building, Building 240,
>   University of Aarhus,
>   8000 Aarhus C,
>   Tel + 45 89 42 1774
>   Fax + 45 8819 6128
>   Email: mikrfdh@svfcd.aau.dk
>
>
>
>
>
>

~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Anna V. Anagnostopoulos, Ph.D.
Division of Biomedical Information Sciences
The Johns Hopkins University School of Medicine
2024 E. Monument Street
Baltimore, MD 21205-2236
tel: (410) 614-3226
fax: (410) 614-0434
e-mail: anna@gdb.org




From tjf at uci.edu Mon Dec 30 18:49:02 1996
From: tjf at uci.edu (Tom Fielder)
Date: Fri Jul 9 14:03:00 2004
Subject: Balb/C knockouts
Message-ID: <v03007802aeed4f41ca8f@[128.200.21.145]>

Does anyone know of a source of CD4 and/or CD8 knockouts in Balb/C
mice? A
colleague of mine has contacted Taconic and Jax and they only have them
in
C57's. Thanks in advance.
Tom

Thomas J. Fielder
UC-Irvine
University Lab Animal Resources
147 BSA
Irvine, CA 92697-1310
e-mail: tjf@uci.edu
phone: 714-824-8579
FAX: 714-824-2003

				
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