I'm preparing to do reverse transcription reactions, but would like each molecule of RNA to be represented once in the cDNA. I was looking for RTs that have RNaseH, but it looks like most RTs have reduced or eliminated RNaseH activity. So, my question is: do RTs generate cDNAs over and over again from the same RNA strand?
Or, does anyone have a recommendation for an RT with RNase H activity?
It is roughly 1:1 reverse transcription. Since you generally cannot quantify cDNA after the RT, you quantify the RNA before (ensure the RNA has good quality and is not degraded over checking 28S and 18S bands on agarosegel) and assume your cDNA has the same concentration afterwards.
This is general procedure for generating cDNA template for qPCR
As others said, one mRNA copy = 1 cDNA copy, otherwise we couldn't do quantitative PCR (qPCR or RT-qPCR) after the RT.
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