Discussion

4 years ago

When should I add the polymerase for hot start PCR?

I keep getting a non-specific band in PCR so I would like to try a hot start PCR manually. How should I prepare the mix and at which step should I be adding the polymerase?

Experiment:

1 Comment


For a hot start PCR you need to prepare the mix without the enzyme. Then at the start of the first annealing step, you should add your polymerase. In this way, your polymerase will not affect any material that annealed on temperature below the one you intended.

Answered 4 years ago


Can you help?

Similar discussions


Start your discussion


Share your thoughts or question with experts in your field

Start discussion
Become shareholder Discussions About us Contact Privacy Terms