Advice needed for RNAse protection Assay

Martin Leach leach at mbcrr.harvard.edu
Fri May 6 08:08:22 EST 1994


> In article <2qbv22$jnr at nntp.ucs.ubc.ca>, Sarven Sabunciyan
> <sarven at ecc.ubc.ca> wrote:
> 
> > Dear netters,
> > 
> > After several years of doing Northerns we have decided to switch to
> > the RNAse protection assay.  I was just wondering about what the in
> > vitro transcription procedures or kits people used to make the RNA
> > probe.  I was also wondering how people isolate their RNA probe.  I
> > am especially curious about if the thickness of the polyacrylamide
> > gel causes a problem with the resolution necessary or the eluting of
> > the probe.  Any sort of helpful hint or favorite procedure would be
> > greatly appreciated.
> > 
We use the method in Current Protocols (by Ed. Ausubel).

Works fine. We use RnaseT1 alone. Dont know what Ambions kits are like,
although their enzymes are cheaper than Pharmacia!! (RnaseT1). Their tech
service is also pretty knowledgeable and have an excellent manual that
comes with their kit. Ask for it. Great trouble-shooting.

Martin
-- 

.....          Martin Leach                Email:leach at mbcrr.harvard.edu 
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