Dig-Northerns

Jaana Tuomainen jtalvine at messi.uku.fi
Mon Sep 11 08:09:51 EST 1995


Request: How to wash off a dig-labeled probe?

We have recently tried in our group the Boehringer 
Dig-kit for Northerns and got some very neat blots 
- with our Southerns it has already been working nicely. 
But the problem here is, that the membrane might be 
more or less "disposable", since at least in Southern 
blots we have not been able to wash off the old probe. 
With plenty of material it is, of course, always possible 
just to run a new gel, but with polyA-blots (which I 
myself have to make) and with some rare samples you just 
cannot spare a new blot for each probing. So, does anyone 
have a good advice for stripping conditions? We use mainly 
the membrane by Boehringer as recommended in the kit 
manual. Also, does anyone have experience in reprobing an 
"old" membrane used in radioactive hybridizations with a dig-
probe? Does it work? 

Or should we just go back to the good old radioactivity-lab?

Lots of thanks for your help!

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