I have some question about qPCR primers. My primers were designed by using Roche universal probe library and one of them(reverse primer) is complementary to the part of sequence of amplicon but forward primer sequence is consistent with the part of amplicon sequence.- why? Do I get double strand cDNA?
My second question is why initial denaturation step occur? I presume that this is needed for denaturation of duplex mRNA and cDNA but I'm not sure.
Ps. sorry for mistake in the topic, I mean qPCR of course
Edited by chaperones, 11 May 2011 - 08:13 AM.