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PCRsheet

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									 PCR Information Sheet                            data input area
                                          Date
                      Name of contact person:
                              e-mail address:
                           Telephone number
                                 Fax number
                                    Affiliation
                               Postal address

                   Gene name (convensional)
               Cytoband in the mouse genome
                            MGI gene symbol*
                        MGI gene ID number*

                        Forward primer Name
                     Amount of forward primer
                     Forward primer sequence 5'
                                                                         Do not put any spaces or hyphens.
                        Reverse primer Name
                     Amount of reverse primer
                     Reverse primer sequence 5'
                                                                         Do not put any spaces or hyphens.
                                           o                               o
      Lower limit of annealing temperature, C                               C
                                           o                               o
      Upper limit of annealing temperature, C                               C

                        Size of PCR product, bp                            bp
Size of coding sequence(s) in PCR product, bp                              bp
        Size of 5' upstream noncoding exon, bp                             bp
      Size of intronic noncoding sequecnes, bp                             bp
     Size of 3' downstream noncoding exon, bp                              bp
                                SNP information      yes, no. or other
                                 Any comments




 PCR Conditions (Overwrite your modifications in the Table below w
PCR reaction mix                                     Standard Conc.                Standard Amount
10 X PCR buffer (with 20mM MgCl2)                         10X                             1ul
additional MgCl2                                            -                              -
dNTP                                                     2.5mM                           0.8ul
Primer F                                                  5uM                            0.4ul
Primer R                                                  5uM                            0.4ul
TaKaRa Ex Taq Hot Start Version                          5u/ul                          0.05ul
template genomic DNA                                    10ng/ul                           1ul
dH2O                                                       --                           6.35ul
                 PCR program                               min                        temperature
                      1                                   4 min                          94C
                      2                                   30 sec                         94C
                     3                                   1 min                     low - high C
                     4                                   1 min                         72C
                     5                                  30 cycle                   go to step 2
                     6                                    4min                         72C
                     7                                    hold                         15C


PCR product sequence information
Please use this expample template. One line holds just 120 letters. (Do not change t

EXAMPLE (This is just an example to show how to provide your information. Please era
cell and put yours.)

tgggcaatgcgcatgcccaatctggacatcctctcttagGGTTCCCGACCACAGTATTGCGGAAAGAGGAATTTTGTCATCCCT
ATGAGTTCTCAAT
forward primer---------Intron 3--------
lyPheProThrThrValLeuArgLysGluGluPheCysHisProLeuLeuLeuLysLysGlyLysIluMetSerSerGlnS

CACAgttcgattcgattactttacgatgcccctgactctttcttctagTCCTGGTTTTTCTGTATTGCGGAAAGAGGAATTTTG
CCACACTACTTAA
erHi---------------Intron 4---------------------
Put the whole sequence of the PCR product including the forward and reverse primer sequences at the beginning and en
Underline the primer sequences.
Put capital (AGCT) and lower-case (agct) letters for coding and noncoding sequences. Namely, use lower-case letters fo
Put amino acid sequence as well as start/stop codons and exon/intron information under the DNA sequence.
            comments if any                                Ver. 080821
            Please fill all the white cells appropriately.
            Also send us an electrophoresis photo(s) of the
            PCR products as a separate electric file(s).




            * for MGI symbol and number, please refer
            http://www.informatics.jax.org/mgihome/no


            Usually 100uM X 200ul.
       3'   5' to 3' direction. Use "AGCT" and "agct" for coding and noncoding sequences, respectively. No other letters in this input. (Please note to use lower-case letter



            Usually 100uM X 200ul.
       3'   5' to 3' direction. Use "AGCT" and "agct" for coding and noncoding sequences, respectively. No other letters in this input. (Please note to use lower-case letter




            Include primer sequences.
            Sizes of coding exons only.



            Particularly between DBA/2 and C57BL/6
            Please provide SNP information if "yes" or
            "other"in the above cell.




e Table below with different color letters.)
       Comment if any




       Comment if any
s. (Do not change the font type or size.)

rmation. Please erase all letters in this


GAGGAATTTTGTCATCCCTTACTACTTAAGAAAGGTAAAATT


IluMetSerSerGlnS

TGCGGAAAGAGGAATTTTGTCATCCCTTACTACTTTTCCCGA

ces at the beginning and ending, respectively.

y, use lower-case letters for noncoding exons (5'- and 3'-UTRs).
 DNA sequence.
ers in this input. (Please note to use lower-case letters for non-coding exons.)




ers in this input. (Please note to use lower-case letters for non-coding exons.)
取り込み   ID   ID2   ID3   プライマー 連携先研究機 連携先責任
                        名称    関名     者
                             0     0
                             0     0
メール   連携先担当       メール2 連携先       遺伝子名       gene symbol primer
      研究員                                               pairL
              0      0       0          0             0
              0      0       0          0             0
primer   予想bp       coding       担当研究員   受取日   配列(5'→ 列 列
pairR                                          3')    1 2
                0            0                      0
                                                    0
bp of primer D2-B6間SNP       PCR結果   singleになったa 最適annealing温   コメント       TGCE条件
                                     nnealing temp. 度

           1 yes, no. or other                                         0
           1
screening状況   担当   位置 向き   Mapping   Accession   memo
                                     No
Tm   memo&文献ま 配列   列1
     たは提供者

								
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