Help: stripping northern blots

denton at usuhsb.usuhs.mil denton at usuhsb.usuhs.mil
Tue Jan 25 18:05:52 EST 1994


Jorges at bcm.tmc.edu (Jorge L. Sepulveda) writes:
>I have been using Zeta probe membranes. The problem is that I can't stripp the 
>probes (boiling in water for 20' or heating overnight at 65¡C in 80% 
>formamide hardly knocks off any counts from the membrane). 
 
In my experience, while this medium gives super high S/N ratios, it
is highly resistant to stripping.  Charged nylon membranes made by
MSI or Schleicher and Shuell give marginally poorer S/N ratios,(IMHO)
but are eminantly more easily stripped of DNA probes than ZETA.
All of these membranes are difficult to strip if you are probing
with riboprobes, presumably as the result of the higher Tm of the
heteroduplex, although there was a recent blurb in one of the
Red Book circulars about using 80% formamide (you refer to this
method, I presume) when stripping the blots.
 
 
My opinions are my own, etc., etc.,
  
R. Rex Denton 



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