Can Taq use RNA as a template?
greg.appleyard at usask.ca
Fri Sep 10 09:07:10 EST 1999
In article <7rb5fb$det$1 at nnrp1.deja.com>,
nicholas_theodorakis at urmc.rochester.edu says...
>The original post didn't mention whether the expected PCR fragment is
>the size expected from a processed mRNA or from genomic DNA, or what the
>source of his RNA is, but note that in many mammalian genomes there are
>sometimes "processed" pseudogenes which have no introns, and would give
>the same size PCR band as one would get from amplifying cDNA. I've
>noticed that I sometimes get this at a low level with beta-actin in
>mouse cells. It's hard to guarantee that your RNA is absolutely free of
>all genomic DNA contamination, especially if you use a "total RNA"
>method such as GITC, Trizol, etc.
The source of RNA is virus. The virus does not have a integrated
pro-viral DNA form and is normally double stranded.
I agree that contamination is a likely possibility for our results but
it is not the only one.
The amplified bands we see are of the expected size.
Tim Fitwater of NeXstar Pharmaceuticals (Gilead Sciences)
wrote an Email reply with this reference:
M. D. Jones and N. S. Foulkes, 1989. Reverse Transcription of mRNA by
Thermus aquaticus DNA polymerase. Nucleic Acids Res. 17(20): 8387-8388.
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