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RT-PCR in a single tube - (Feb/15/2012 )

Dear Forum members,

Starting from purified 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?


There are many. Just google "one-step RT-PCR".
Life Technologies
and so on..


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.

-Rachel Ruhlen-