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?
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.
More information about the Methods