mRNA storage

Tracy Aquilla aquilla at salus.med.uvm.edu
Thu Sep 1 11:38:05 EST 1994


In Article <9408291849.AA0192 at rnaworld.bio.ukans.edu>,
pgegen at RNAWORLD.BIO.UKANS.EDU wrote:
>Question was: How do you ensure that DEPC is removed?
>
>Answer -- I have no good answer! DEPC should hydrolyze, upon
>heating, to CO2 + H2O. Simplistically, I'd say if you can
>still smell the DEPC (fruity, ester-like odor) then it's not all hydrolyzed. 
>The few times I tried to treat water with DEPC, I autoclaved it twice but I 
>could still smell the DEPC. Since none of my transcription reactions worked 
>with that water, I gave up on DEPC.
>
>o-----------------------------------------------------------------------------o
>| Peter Gegenheimer                          |  pgegen at kuhub.cc.ukans.edu     |
>| Departments of Biochemistry and of Botany  |  voice: 913-864-3939           |
>| University of Kansas                       |  FAX  : 913-864-5321           |
>| 2045 Haworth Hall                          | "The sleep of reason produces  |
>| Lawrence  KS  66045-2106                   |  monsters."              Goya  |
>o____________________________________________|________________________________o
>

Actually, DEPC (diethylpyrocarbonate) should hydrolyze into CO2 and EtOH
when added to water. While it is true DEPC-treated water usually has some
residual odor, I think it is mostly from the ethanol, not DEPC. If you are
worried about it, you should autoclave the water and then shake it
vigorously for a few hours. Repeated autoclaving will not get rid of the
smell. If you have a good clean water supply, i.e. a Milli-Q system that is
maintained well, I would skip the DEPC altogether, but make sure the
glassware is RNAse-free. DEPC is notorious for degrading RNA as well as
inhibiting many enzyme-catalyzed reactions, so if you can, you should avoid
using it. Sometimes though, it is really needed.



Tracy Aquilla, Ph.D.
Molecular Physiology and Biophysics
University of Vermont
aquilla at salus.med.uvm.edu



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