TE Buffer for RNA

Andrew Doherty A.Doherty at Bris.ac.uk
Fri Nov 29 03:59:06 EST 1996


brett wrote:
> 
> >Help!
> >In making TE buffer for RNA use is it common practice to treat freshly =
> >made up TE with DEPC?  I've heard that Tris interacts with DEPC =
> >unfavorably therby negating the RNase-inactiving effects of the latter - =
> >although I've never seen this documented.  If this is true, can someone =
> >suggest how RNase-free TE can be made?
> >Many thanks!
> >
> >Karl Franek
> >Microbiology & Immunology
> >LSU Medical Center
> >Shreveport, LA 71130
> >
> >kfrane at mail.sh.lsumc.edu
> 
> Make it up in RNase-free water. We have found that our freshly squeezed milli-Q
> water has undetectable RNase activity; you may want to DEPC-treat your supply,
> as you wish. Anyways, we then make up a 1M Tris stock and a 0.5M EDTA stock from
> bottles of reagent reserved for this, weighed onto baked pieces of foil or
> directly into baked glassware or new 50ml Falcons (they seem to come
> RNase-free). Do not use spatulas, stirs bars, or pH probes, although
> individually wrapped, sterile plastic pipettes can usually be trusted.
> Adjust pH by adding the right amount of acid/base based on titration of a
> small
> aliquot. I use these same approaches to all my RNA work, and generally shun
> DEPC. Haven't had a problem (yet!)
> 
> Brett Lindenbach
> 
> Program in Immunology
> Washington University - St Louis
> brett at borcim.wustl.edu
Couln't agree more - I never use DEPC for RNA work, just keep everything
separate, NEVER put a spatula in a pot of anything used for RNA and
generally keep everything VERY clean and you'll be OK
-- 
*************************************************************
Dr Andrew Doherty		email -  a.doherty at bris.ac.uk
Dept. Anatomy			Tel (0117)9287421
School of Medical Sciences	Fax (0117)9287402
University of Bristol
University Walk
Bristol UK
BS8 1TD
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