Annealing temperature for PCR
Posted 10 August 2004 - 07:43 PM
Posted 10 August 2004 - 09:22 PM
Even if i subtract 5-10 from tm...then should I subtract from tm of forward or reverse primer?
Posted 10 August 2004 - 09:36 PM
normally I'd subtract from the Tm of smaller primer. If you are worried about the specificity of your reaction you can do a test run with a few different annelaing temperatures