northern blots

wetsel_r at wums.wustl.edu wetsel_r at wums.wustl.edu
Mon Nov 15 13:05:49 EST 1993


In a previous article, helgew at LJCRF.EDU (Helge Weissig) wrote:
> 
>How do you strip your blots? I have problems with very "sticky" probes even
>after stripping twice w/ boiling 0.1% SDS (membrane is Hybond-N, Amersham)?
> 

Yes, Hybond-N+ from Amersham, and yes again - boiling in basically water 
using as little SDS as possible 0.05%...  I've had to boil them as many as 
four times to get real sticky probes off.  Even after this protocol, we'll 
often expose overnight just to know what kind of "carry-over" we're going 
to get, then go ahead and reprobe.  Sometime's you'll wind up with one of 
those blots where the probe will never come off no matter what you do and 
the best course of action there is to run the bloody gel over!

I've found 0.1M NaOH works real well for reprobing Southerns... I once 
'brain cramped' and did this to a northern.  It goes without saying that I 
had to run the northern again when the positive control didn't even come 
up!  ;-)

David
haviland at kids.wustl.edu



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