RT-PCR

Johanna Fehling jf510 at york.ac.uk
Wed Aug 3 10:41:16 EST 2005


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

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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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