Reverse-transcription - how much RNA will be converted into cDNA (Apr/09/2007 )
I have been getting more than 200ug cDNA (single stranded) from 1ug total RNA in RT reactions. I am confused because I would assume the amount of cDNA would be less than the starting RNA (will probably be twice the amount of the RNA base on the OD reading if RNase H is not used afterward). Or, is there a guildeline for the yield of cDNA in RT? I would definitely appreciate it if anyone can help me with this. Many thanks!
what primer are you using for RT?
I have used both random hexamer and oligo dT
I think that with random hexamers you can get binding to multiple places on the RNA then because there is helicase activity of the RT you can make more than one copy of each sequence?? Maybe this is why you get more cDNA than 1:1...
How did you quantify your cDNA?