DEPC treatment of buffers
Zeiler, Brian M'bio
BrianZ at MICROBIO.LIFESCI.UCLA.EDU
Thu Jun 15 20:03:40 EST 1995
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From: BIOSCI-REQUEST
To: methods-and-reagents
Subject: RE: DEPC treatment of buffers
Date: Thursday, June 15, 1995 10:48PM
>I know you can't DEPC-treat Tris buffers, but does anyone know if there are
>common buffers that you can treat? (eg MOPS, HEPES in particular). Or is
it
>always best just to make them up with DEPC-treated autoclaved water?
>Any advice greatly appreciated,
Neil Saunders.
:nsaunders at molbiol.ox.ac.uk
:best advice: do not use DEPC at all.
:If you do not use tap water and include placental
:ribonuclease inhibitor RNA must survive.
:It is very difficult to inactivate 100% of DEPC.
:Andrei
Whoa! Not true! Placental RNAse inhibitor DOES NOT work against all
RNases, just read the spec sheet, it even includes RNases that it is known
to NOT work against. We autoclave our DEPC solutions and have never had a
problem with any application.
Sorry I did not answer your question about buffers but I did not want anyone
in netland to lose their precious RNA. I use DEPC-treated autoclaved water
or milliQ water to make up HEPES buffer and that works fine, just be certain
that your HEPES is dedicated for RNA use and is handled by properly trained
personnel.
Brian
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