Description: Hot start Taq DNA Polymerase is a recombinant Taq DNA polymerase mixed with its specific antibody that inhibits polymerase activity at ambient temperatures. This prevents extension of nonspecifically annealed primers and primer-dimers formed at low temperatures during PCR setup and the initial PCR cycle.
Description
Hot start Taq DNA Polymerase is a recombinant Taq DNA polymerase mixed with its specific antibody that inhibits polymerase activity at ambient temperatures. Due to specific binding of the inhibitor, the Taq DNA Polymerase is provided in an inactive form. This prevents extension of nonspecifically annealed primers and primer-dimers formed at low temperatures during PCR setup and the initial PCR cycle. The DNA polymerase is activated in a temperature-dependent manner (at 95ºC) during the start of PCR. Once dissociated from the inhibitor, the Taq DNA polymerase regains its activity. This makes Hot Taq DNA Polymerase ideal for quantitative real-time PCR.
Package
Taq DNA polymerase (5U/μl) Cat No. RE-12-002 200 U
10X Buffer(Mg2+ free) 1000 μl
100 mM MgCL2 100 μl
Recommend PCR Protocol
Note: (1) The hot start Taq DNA polymerase used in this format is able to tolerate higher temperature up to 98℃ so that customer can use denature temperature up to 96℃ with denature time as short as 10 seconds in his PCR cycling.
(2) This format of DNA polymerase is of higher fidelity than conventional Taq.
Model Protocol:
In a 0.2ml thin-wall PCR tube, add：
Taq DNA polymerase 1μl
10×buffer 5μl
Primer 1 2μl
Primer 2 2μl
Template 1～10μl
Add H2O to 50μl
Spin briefly before cycling at the following conditions:
Predenature: 95℃×15min
Cycling: 94℃×10s, (Tm-5)℃×20s, 72℃(fragment length in kb/2×60)s, 30～40 cycles
Post Extension: 72℃×3min
After finished, spin at high speed for not less than 2mins, then load onto a agarose gel with proper concentration.
Storage
Store all components at -20℃(non-frost-free). Thaw 10X Buffer at room temperature just prior to use and refreeze immediately.
