What can my annealing temperature be if both my forward & reverse primer have the same Tm according to synthesis report?

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Syed A Ali

It can be 55-57C i.e., 3-5C below the calculated Tm. However, the optimal annealing temperature should be determined emperically.

Chinwe Juliana Iwu

Hi Mr Afolabi, I agree with Syed. You optimise using the temperature ranges provided above. Also, having a positive control for the organism makes optimisation a lot easier.