Primers with 2 different annealing temp - (Jan/27/2015 )

Hi there all

I have a primer with two different annealing temperature.

The forward is 60.7C and reverse 58.8 C, please advice how to go about this

regards

That is quite usual. You want them as close as possible, but successful primer pairs can even have differences of 20'C.

Do the PCR somewhere in the range of 55-60 C.