Now I have Using Q5® High-Fidelity DNA Polymerase (M0491) Thermocycling Conditions for Routine PCR itis recommended Initial Denaturation 98c? But I don't have any results?!! Also, I try another Taq polymerase, Addbio (AddStart Taq DNA Polymerase) Product Code 18101X; it has a positive PCR band. I can change the initial denaturation to 94 c, like the picture attached.
I can use Q5® High-Fidelity DNA Polymerase (M0491) but at 94 C?
Paul Rutland
Q5 uses an aptomer to block the active site of the enzyme and this must be removed before there can be any polymerase activity. The company says that 98c is needed to remove this aptamer so I would not change the 98c step but look elsewhere ( maybe annealing temperature of the primers) for a result to your problem
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