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RT-PCR in a single tube


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3 replies to this topic

#1 Chelo

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Posted 15 February 2012 - 08:30 AM

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!

#2 Curtis

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Posted 15 February 2012 - 09:33 AM

why not use ordinary reverse transcriptase?

#3 Trof

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Excellent

Posted 16 February 2012 - 08:15 AM

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

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#4 Rachel Ruhlen

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Posted 16 February 2012 - 08:21 AM

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.




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