DNA Polymerase for Hot Start PCR by alj19178


									                               SurePRIME                                                     ™

                 DNA Polymerase for Hot Start PCR
SurePRIME™ DNA Polymerase is a highly purified form of recombinant Taq DNA Polymerase
that has been chemically modified by the addition of heat-labile blocking groups to specific
amino acid residues. Prior to PCR, in its inactive state, SurePRIME™ DNA Polymerase is inca-
pable of extending primer-dimers or mis-annealed primer-template species that form below the
specific annealing temperature. The 95°C incubation step therefore serves to activate the
enzyme and also to ensure a completely "clean" initial PCR cycle.

Easy-to-use                                                                                          Highly specific
Once activated SurePRIME™ DNA Polymerase is functionally equivalent to Taq DNA
                                                                                                   DNA amplification
Polymerase, and so the only PCR program modification required is the addition of a 15 min.
pre-incubation step at 95°C. Efficiency can be improved by progressive activation (initial acti-
vation 5 min. at 95°C followed by 1 min. activation at 95°C at each cycle).
                                                                                                    Low background
Performance under challenging PCR conditions
A comparison of SurePRIME™ DNA Polymerase and classical recombinant Taq DNA                            Higher Yields
Polymerase was performed under challenging PCR conditions. A 400 bp region of the human
ß-globin gene was amplified using specially designed non-optimized primers. The primer set
contained mismatched bases at the 5' end to introduce restriction sites.
 Taq DNA       SurePRIME™      To further influence the conditions for non specific priming,
Polymerase    DNA Polymerase
                               the PCR reactions, once set up, were incubated for 30 min at
                               25°C. During set up, the residual activity of Taq DNA
                               Polymerase elongated mispaired primers leading to non spe-                Saves time
                               cific PCR products. On the contrary, SurePRIME™ DNA
                               Polymerase exhibited no residual activity, and thus did not
                                                                                                          and effort
                               elongate mispaired primers.

                               Comparing Taq and SurePRIME™ DNA Polymerases
                               In this experiment amplification conditions favoring the
                               formation of non-specific product were purposely employed
                               – non-optimized primers and 30 minutes pre-incubation at
                               room temperature.

                    For more information visit our website at www.qbiogene.com
SurePRIME™                                      DNA Polymerase for Hot Start PCR

Key Features                                                        Cat #         Description                                      Quantity
1) SurePRIME™ DNA Polymerase exhibits no activity at                EPHSP025 SurePRIME™ DNA Polymerase                                250 U
   room temperature. This means that variations in the              EPHSP525 SurePRIME™ DNA Polymerase                            5 x 250 U
   time taken to set up reactions will not affect repro-          A 10X incubation buffer without MgCl2 is provided
   ducibility of results.                                         A solution of 25 mM MgCl2 is also supplied

2) SurePRIME™ DNA Polymerase provides highly specific
   DNA amplification even when using non-optimized

SurePRIME™ Also Offered In These Convenient Formats:
Available in CORE kits                                            Two types of kits are supplied
                                                                    Cat #                     Units*                     Qty of dNTPs Mix
SurePRIME™ CORE kits contain all the reagents (presented
as separate items) required for hot start PCR: SurePRIME™           SurePRIME™ CORE Kit 10 (with dNTPs at 10mM each)
DNA Polymerase, incubation buffer                                   EPHSK105            5 x 250 U              15 µmol each dNTP
and high purity dNTPs Mix (>99%).                                   SurePRIME™ CORE Kit 25 (with dNTPs at 25mM each)
                                                                    EPHSK255            5 x 250 U              15 µmol each dNTP
                                                                  *Units of SurePRIME™ DNA Polymerase

Available in a Mastermix                                            Cat#             Description             Reactions (Units)*          Size
SurePRIME-&GO™ Mastermix contains all the                           EPHAG100         SurePRIME-&GO™               100 (100 U)             1 ml
components required for hot start PCR with                          EPHAG110         SurePRIME-&GO™             1,000 (1,000 U)      10 x 1 ml
the exception of template and primers.                            *Units of SurePRIME™ DNA Polymerase
SurePRIME-&GO™ Mastermix is provided
5X concentrated and is designed to provide                        SurePRIME-&GO™ 1 x C composition:
1 U of SurePRIME™ DNA Polymerase per                              SurePRIME™ DNA Pol   0.02 U/µl
                                                                  dNTPs each            100 µM
50 µl reaction.                                                   Tris-HCl pH8.3         10 mM
                                                                  KCl                    50 mM
                                                                  MgCl2                 2.5 mM
SurePRIME-&GO™ Mastermix is stable for over one year              Glycerol                 1.8%
at +4°C.


              NORTH AMERICA                                UK
              Phone : 1-800-424-6101 or (760) 929-1700     Phone : 0800 328 8401
              E-mail : order@qbiogene.com                  E-mail : ukorders@qbiogene.com
              FRANCE                                       GERMANY
              Phone : 03 88 67 54 25                       Phone : 0800 409 0580
              E-mail : frorders@qbiogene.com               E-mail : bestellungen@qbiogene.com

To top