Storage of RNA
hamel at cc.umanitoba.ca
Thu Oct 13 14:27:21 EST 1994
RNA should store well in sterile dH2O in freezer.
We have MANY aliquots of total RNAs obtained from several types of
specimens (feces, tissues, whole blood, etc, etc) ... ALL store WELL in
-20*C PLUS used daily (frozen-thawed MANY times) ... used for RT-PCR ...
ALL are still okay, right down to last bit used (most aliquots are 50 uL,
with 1 uL used per reaction).
If stored in formamide (such as used for northerns) ... simply ppt with
alcohol and NaAc ... add NaAc to 0.3 M final conc and use 1-2 vol of
isoprop or 2-3 vol of EtOH, mix well, incubate -20*C for at least 1 hr
before harvesting pellet ... formamide will be washed away (remains in
supernatant along with the alcohol).
Andre Hamel email: hamel at cc.umanitoba.ca
Manitoba Government tel: 204/945-7630
Department of Agriculture FAX: 204/945-8062
Veterinary Services Branch
Winnipeg, Manitoba, CANADA
In article <sticknbd-121094224623 at 188.8.131.52> sticknbd at miranda.cc.vanderbilt.edu (B Stickney) writes:
>In article <1994Oct7.192254.1 at molbiol.ox.ac.uk>, rhubner at molbiol.ox.ac.uk
> > In article <ros.2.2E911FE4 at molbiol.uct.ac.za>, ros at molbiol.uct.ac.za
--- stuff deleted ---
> > > I have been storing RNA samples from both E.coli and Thiobacillus
>> > ferrooxidans in DEPC water or RNA storage buffer, 20mM sodium phosphate pH
>> > 6.5 1mM EDTA. The T.ferrooxidans appears to be ok on a agarose gel, but no
>> > longer gives good results on a Northern Blot, and I presume this is due to
>> > degradation of the RNA. I store the RNA at -70 C and thaw and refreeze it
>> > every time I use it, so this could also be causing problems. I see some
>> > people store their RNA in formamide at -20 C, how do they then get rid of
>> > the formamide, when they want to do primer extension for example?
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