What can be the appropriate annealing temp for PCR if tm of forward primer is 57.9 and that of reverse primer is 60?

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Nadine A. Gund

Hi, the annealing temperature should be 54-56 °C (you can do a gradient PCR). Cheers, Nadine

Sibtain Afzal

Hi Rene

You may do the touchdown pcr. You will get your required results definetely.

Best Regards

Ananda Ayyappan Jaguva Vasudevan

I would average it so it is 59 degrees, and Annelaing tem=Tm-5deg; so I would try from 54degrees.

Md Suhail Alam

I would go for gradient PCR from 52-58 with a increment of 1degree.