A successful reverse transcription reaction - (Oct/12/2006 )
I want to know what precuations must be taken so as to get a perfect result after reverse transcription of total RNA, the RT enzyme which I am using is active at 37 degree C.Ofcourse my aim is to reverse transcribe mRNA populations from total RNA. I don't want that folded sec. structure of mRNA cause any hinderence in this step.
Well, you could use a thermo-cycler with a program of repetitive changing the temp to, say, 45-50 centigrades just for 30 sec (denaturation) and 10 minutes of 37"C (enzyme activity).