Single or double-stranded cDNA?
Posted 19 March 2011 - 11:48 AM
I posted this question elsewhere but nobody answered. It is stupid basic question but I need to know.
I do two-step RT-PCR, i.e. first I do the RT reaction, and then - basic PCR.
I use recombinant RevertAid M-MuLV Reverse transcriptase from Fermentas - in datasheet they say that the enzyme has RNA- and DNA-dependent polymerase activity and an RNase H activity specific to RNA in RNA-DNA hybrids. As I imagine it is something like 3 in 1 - RNase activity cleaves the RNA, and DNA-dependent polymerase builds the second strand.
Thus, I think that in the end of reaction I have double stranded cDNA.
It that wrong?
Posted 19 March 2011 - 02:10 PM
Posted 20 March 2011 - 04:28 AM
Thank you very much.
Now I understand it.