Quantitative RT-PCR

mbarnhart at my-dejanews.com mbarnhart at my-dejanews.com
Thu Jun 18 09:58:58 EST 1998


We are developing competitive RT-PCR for three genes and plan to work with
more in the future.  Since we typically only use about 20% of the RT
reaction, I was wondering if we could use multiple competitors in a given
reverse transcription so that one reaction could be used for multiple genes. 
For example:

    RT rxn 1:  Sample RNA + 10 e10 copies Actin competitor + 10 e10 copies of
myosin competitor

    RT rxn 2:  Sample RNA + 10 e9 copies Actin competitor + 10 e9 copies of
myosin competitor

We would be using Random primers in the RT reactions, so presumably first
strand cDNA would be made for all the RNAs.

I can't think of any reason why this shouldn't work, but I hoping to get some
input and maybe find out if anyone else has tried this.

Michael Barnhart
Laboratory Supervisor
Biology Department
University of Houston

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