Stripping riboprobe northerns

Aaron Pawlyk aaron at ccat.sas.upenn.edu
Tue Sep 5 15:22:38 EST 1995


Eugene Chomey (echomey at gpu3.srv.ualberta.ca) wrote:
: I am having problems taking off my riboprobes on northern blots.
: Does anyone have any proven methods for their efficient removal?
: Advice appreciated.

I don't have any good advice for this.  I have never been able to get
riboprobes off of my Northern filters.  Even pouring boiling 0.1% SDS on
them and letting them sit for 3 minutes doesn't get it all off.

My suggestion would to use DNA probes.  A great way to get high activity
DNA probes is to PCR amplify up some probes with a radioactive nucleotide
in the mix!  Should give you the sensitivity you're looking for (if you're
just using riboprobes for the heck of it, just use random priming).




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