RNA PURIFICATION

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:
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>From: sgsmith at tcd.ie (Stephen Smith)
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>Subject: re:RNA PURIFICATION
>Date: 17 Aug 1996 06:55:11 -0700
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>Hello AJ, 
>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 
>(Promega)).

>All the best,
>Stephen.

>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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