non-radioactive RNAse protection assays

Jeffrey D. Saffer jd_saffer at pnl.gov
Tue Nov 29 10:46:40 EST 1994


In article <39rfvn$aq0 at inxs.ncren.net>, Patrice Ferriola
<Ferriola at ciit.org> wrote:

> Does anyone have any ideas about non-radioactive detection methods
> for RNAse protection assays?  I presently use 33P or 35S-lableled
> riboprobes, but would like to make a probe with longer shelf life.
> Is there any way to label the riboprobe with fluorescent nucleotides and
then detect the protected/fluorescent bands directly on a urea gel?  I
don't want to transfer the gel.  Applied Biosystems couldn't
> provide any answers.  Any ideas?
> 
> thanks

My lab has been looking also.  As far as we can tell, there are no good
options available right now that don't include a transfer step.  Molecular
probes now sells a fluorescent-UTP, but it apparently has some drawbacks
(e.g., poor incorporation, sensitivity to the RNAse, etc).  If you find an
alternative, I'd like to know.



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