Taek H. You
tyou at postbox.acs.ohio-state.edu
Sat Aug 17 14:09:22 EST 1996
In article <3215DFF8.B2A at tcd.ie> sgsmith at tcd.ie (Stephen Smith) writes:
>From: sgsmith at tcd.ie (Stephen Smith)
>Subject: re:RNA PURIFICATION
>Date: 17 Aug 1996 06:55:11 -0700
>Organization: BIOSCI International Newsgroups for Molecular Biology
>Sender: daemon at net.bio.net
>Message-ID: <3215DFF8.B2A at tcd.ie>
>Reply-To: Moyne at vax1.tcd.ie, Institute at vax1.tcd.ie, Trinity at vax1.tcd.ie,
> College at vax1.tcd.ie
>With regard to removing contaminating DNA from RNA samples, have you
>tried selective precipitation of the RNA?
>Add an equal vol. of 5 M Ammonium Acetate to sample, incubate on ice
>20 min, pellet RNA by centrifugation @ 10,000 x g @ 4 degrees for 10
>min. Wash pellet with 70% EtOH, dry briefly and resuspend in RNase
>free water (you may wish to add RNase inhibitor such as RNasin
>All the best,
>ps Your DNase might not be working due to lack of (or chelation of)
>Mg2+/Ca2+ in reaction mix.
Also, look at the Epicentre technical brochure (latest issue).
It has a technical tip about selectively precipitating RNAs.
Very simple and quick. They claimed recovery is almost 100%.
Sorry that I don't have the protocol handy right now.
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