RNase Protection Assay-HELP!

Vesselina Ivanova vivanova at iastate.edu
Tue May 13 21:46:40 EST 1997

Hello, everybody!

Does anyone have any experience with RNase Protection Assay? I have been
doing it for a few months now, using a non-radioactive method for
detection (biotin) and I have problems with transferring from the PAGE
to a nylon membrane (low efficiency of transfer). My other problem is
that after I stain the urea/PAGE with EtBr I see the same band pattern
with my control probe (beta-actin) and with my probe (in this case for
titin). It's absolutely the same picture-with actin, with titin and with
both probes. What I am looking for is differential expression of titin
at different stages of primary chick embryo muscle culture and if there
is no way of comparing titin's expression with that of actin
(housekeeping gene), I am in a bad shape. I would greatly appreciate any help!

Thanks a lot in advance!

Vesselina Ivanova

Graduate student in the Department
of Biochemistry and Biophysics
Iowa State University
Vesselina Ivanova
vivanova at iastate.edu

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