denature RNA and miRNA Q-PCR - (Feb/04/2009 )
I prepared my total RNA using mirVANA miRNA isolation kit. Then I heated the RNA @58 degrees for 10min and store the sample in -80 freezer. Now i'm going to perform a Taqman miRNA Q-PCR. When i read their protocol, they suggested not to denature the RNA because it might reduce the yield of cDNA for some miRNA target. Has anyone used denatured RNA for Taqman miRNA assay? Would the denaturation step be a big problem?
I remembered the protocol saying do not denature RT-primers for miRNA when prepare cDNA.