i extracted RNA from human cell line.and converted it in to cDNA (using two steps TR Kit.
now iam goning to do conventional PCR reaction for detection specific gene expression.
so iam asking about:
do i need to convert cDNA in to double strand before pcr ?
or immediatly run the pcr reaction with single stranded cDNA?
what is the time needed for the pre-denaturation step for cDNA in conventional PCR?
and is it differ according to the source of RNA?
and if it is a single strand cDNA , i think that the pre-denaturation step in negligble or what??
Edited by aidaelshaer, 01 October 2010 - 03:44 PM.