jph5 at leicester.ac.uk
Wed Aug 24 12:09:31 EST 1994
I am having great difficulty in maintaining stocks of "viable",
(i.e RT-PCRable), viral mRNA, and would be very grateful of any suggestions.
Initially, I extract viral positive sense RNA using RNAzol, (Biogenesis), and
store the product in 50 microlitres TE buffer with 0.25 microlitres RNasin,
(Promega). The components for the TE buffer are individually produced as stock
solutions, autoclaved, and mixed when required. The RNA is stored in DEPC
treated, sterilised, Eppendorfs, (though the glassware in which the TE buffer
is stored and the water used in the TE buffer components are not DEPC treated).
RT-PCR on freshly isolated RNA works well, but after 2 days at -20oC
the RNA seems to have disappeared!
Thanks for any advice!
jph5 at le.ac.uk
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