General PCRTransgenomic T-Taq DNA polymerase is a recombinant, thermostable, 94-kDa DNA polymerase cloned from Thermus aquaticus in Escherichia coli. The enzyme possesses 5'->3' DNA polymerase activity and 5'->3' DNA exonuclease activity, but not 3'->5' DNA exonuclease activity. T-Taq is > 95% homogeneous by SDS-PAGE. The enzyme generates PCR products with 3'-dA overhangs. - T-Taq has been developed with a 10x buffer.
- T-Taq is used particularly for colony PCR win conjunction with the SURVEYOR Check-It™ Kit.
T-Taq Kits and dNTPs Cycling ParametersThe optimal conditions for amplification will vary depending upon the primers and the thermal cycler used. Typical cycling parameters are: | | 94°C | 2 min X 1 cycle | | | 94°C | 30 sec | | | Ta°C | 30 sec X 25-30 cycles | | | 72°C | 1 min per 1kb | | | 72°C | 5 min | | | 4°C | Hold |
Calculating Annealing TemperatureDetermine the annealing temperature (Ta) by calculating the Tm for each primer using the following equation: Tm = 63.728 + (0.41 x %GC) – (600/length)
where %GC = percentage GC of the primer
and length = length of the primer in nucleotides Ta is the average of the two primer Tms +3 °C
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