RT-PCR in a single tube - (Feb/15/2012 )
Dear Forum members,
Starting from purified RNA...is it possible to perform a retrotranscription followed by a PCR amplification in a single tube? What I mean is whether it is possible to have a mix of a retrotranscriptase and a Taq-polymerase in a single buffer compatible with both enzymes and to run the RT program at -let´s say- 37ºC and to follow with the PCR program.
If the answer were "yes", I would appreciate any references or commercial product names.
Thank you all in advance for your kind answers!
why not use ordinary reverse transcriptase?
I have used SuperScript III One-Step from Invitrogen with success. It's really easy and I've never had any trouble with it, although my RT-PCR experience isn't as extensive as many people's.