RT PCR-any suggestions?

Annette C. Hollmann ah690549 at mbcr.bcm.tmc.edu
Thu Aug 4 12:37:41 EST 1994


In article <aquilla.1126332364C at sadye.emba.uvm.edu> aquilla at salus.med.uvm.edu (Tracy Aquilla) writes:
>Hi World,
>    I am thinking about using RT PCR to clone several cDNAs, using total RNA
>as template. I am wondering if anyone who has used this technique has any
>suggestions for me. Also, I have heard about a new enzyme sold by
>Perkin-Elmer Applied BioSystems division which has both reverse
>transcriptase activity as well as DNA polymerase activity. I think this
>sounds like a good thing, but again, I would like to hear from anyone who
>has used this enzyme. Any suggestions are welcome. Thanks for the help!
>    Tracy

Perkin-Elmer sells two kinds of RT-PCR kits. One has the all-in-one RT/PCR
enzyme and one has regular RTase and regular Taq. The kit with the all-in-one
enzyme is not recommended for sequencing because the products are not pure
enough. It might be ok for cloning but I would stick with the separate RTase
and Taq. I have used the GeneAmp RNA PCR kit and it works okay once it is
optimized.

Annette
ah690549 at mbcr.bcm.tmc





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