I have used a Taq Polymerase with less Proof reading and fidelity. I have a doubt that some mutation may be occurred during the PCR. One drawback is the polymerase doesn't have 3'-5' exonuclease activity.
Can anyone tell me the method to prove that mutation happened due to the Taq Polymerase only and not from the mother DNA.
Repeat the PCR a couple of times from the parent DNA and sequence all the different amplicons. If the same mutation is present in all the amplifications, then it is possible that it was from the parent template, else because of Taq.
Another way is to use a proof-reading enzyme on the parent DNA, and compare the sequences of both.
Taq polymerase is making errors with a frequency around 1 base to every 3.300-3.700, which is a lot (200-fold) if you compare it with a Hi-Fi which makes 1 every 700.000-750.000.
Cheers
I agree with Devi prasad. Comparing the PCR amplicons of your Taq and a high fidelity polymerase will let you know the answer. If the same mutation happens in both cases, it is on your DNA template.
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