mRNA storage

Tracy Aquilla aquilla at salus.med.uvm.edu
Tue Aug 30 09:49:29 EST 1994


In Article <33fusb$31q at mserv1.dl.ac.uk>, "J.P. Hays" <jph5 at leicester.ac.uk>
wrote:
>Dear Colleagues,
>
>  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!
>
>
>John Hays
>
>jph5 at le.ac.uk
>

I have stored RNA samples in plain distilled water and/or TE for over a year
at -80, with multiple freeze-thaw cycles and no degradation problems. I do
not suggest adding RNAsin. Presently, I am using a product called FORMazol,
from Molecular Research Center, which is just 'stabilized' formamide. This
solvent creates a denaturing environment, and I can store the samples at
-20, which is more convenient. I have stores RNA for about a year this way
with no problems.

Tracy Aquilla, Ph.D.
Molecular Physiology and Biophysics
University of Vermont
aquilla at salus.med.uvm.edu



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