RNase Protection Assay; DEPC for everything?

David L. Haviland, Ph.D. dhavilan at IMM2.IMM.UTH.TMC.EDU
Thu Feb 26 17:44:02 EST 1998

At 16:43 2/25/98 -0800, Rolands G. Aravindan wrote:


>I apologize for troubling the newsgroup, and specifically you, David.

Don't apologize, this is what this newsgroup is for...

>While investigating setting up RPA, I was told that I should use DEPC
>treated water for every reagent as a general principle. Is this OK? Any
>comment would be appreciated.

Absolutely!  I would refer you to Maniatis or the Harvard manual (Curr.
Protocols in Molec. Biology).  In short, all of your reagents for RNA
should  be made up and "deemed" RNAse-free, only to be touched with gloved
hands and sterile pipets.   My lab has even bought separate reagents
strictly for RNA work.   Common reagents and  RNA reagents are never
intermixed and are in separate places.  All your reagents used for RNA work
should be made with DEPC-treated water.

And yes, per the one comment, the DEPC mixed with water, then autoclaved.
This DEPC treated water is then used to make all necessary solutions.

Hope this helps,
 David L. Haviland, Ph.D.
 Asst. Prof. Immunology 
 University of Texas - Houston, H.S.C.
 Institute of Molecular Medicine  
 2121 W. Holcombe Blvd.  
 Houston, TX  77030 
 Internet:"dhavilan at imm2.imm.uth.tmc.edu" 
 Voice: 713.500.2413  FAX: 713.500.2424
Never take life seriously. Nobody gets out alive anyway.

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