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RT-PCR

RT-PCR - Protocols and Methods Forum

RT-PCR - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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Old 08-03-2005, 03:41 PM
Johanna Fehling
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Default RT-PCR



Does anyone know of an RT-PCR kit or protocol that would select for
long polyA tails? I am trying to do RT-PCR on protozoan cell cultures
contaminated with a mix of bacteria. Getting rid of the bacteria
doesn't seem to be an option (many things have been tried). Although
some bacterial transcripts are polyA'd, I think these tend to be much
shorter than in euk transcripts. I've been using the Strategene kit
"StrataScript Two-tube RT-PCR system with PfuUltra" but still seem to
be getting a lot of bacterial genes in my PCR products. I know polyU is
supposed to be more selective than polydT. Does anyone know of any kits
that use polyU, or any other tricks I might try?

Cheers,
Johanna

--------------------------------------------------------
Johanna Fehling
Postdoctoral Research Fellow
Department of Biology, Area 3
University of York
PO Box 373
York, YO10 5YW
United Kingdom

phone#: ++44 (0)1904 328633
fax#: ++44 (0)1904 328505
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