Does the activation of AmpliTaq Gold™ at 95°C for 10 minutes interfere with the half-life of the enzyme?
The half-life of AmpliTaq Gold™ at 95°C is 40 minutes. This is with constant incubation at the described temperature. It is a known fact that during PCR, the sample, and therefore the enzyme, is truly only incubated at the programmed temperature for approximately 20 seconds. Therefore, the cycling half-life of AmpliTaq Gold at 95°C is approximately 100 cycles. Example: AmpliTaq® DNA Polymerase experiences about 20 seconds (0.33 min) at 95°C per PCR cycle. t1/2 is at least 33 minutes; (35-40 mins) therefore, 33 min / 0.33 min/cycle = 100 cycles 100 PCR cycles reduces enzyme activity by 50%.
