stripping Northerns

Bill McKendree mckendre at ASRR.ARSUSDA.GOV
Mon Nov 10 08:49:56 EST 1997


I routinely strip and re-probe Northerns with biotin labeled probes.  
They tend to get noisy  after three rounds of this, however.  The only 
problem I've had with the biotin labeling system is with consistency in 
probe labeling.

Bill Mckendree
USDA-ARS
Orlando, FL

On Fri, 7 Nov 1997, Lely Chow wrote:

> I have never been able to successfully strip Northern probed with DIG
> labelled probes.  It seems to be a problem intrinsic to using DIG probes,
> as I do not have the problem when probing with p32 probes.  I would
> suggest switching back to p32 probes.
> 
> 
> Lely Chow
> Department of Biological Structure
> University of Washington
> Seattle, WA 
> 
> 
> On 29 Oct 1997, Dianne Emslie wrote:
> 
> > 
> > I am having problems stripping Northern blots probed with DIG labelled
> > oligos and hope someone may have a better method.  I have tried boiling in
> > 0.1% SDS which seems to remove the chemiluminescent substrate but not the
> > oligo and I have tried the DIG Northern probe-stripping solution which
> > contains 50% fomamide. That seemed to strip the probe off but also the
> > RNA.  Does anyone have any suggestions? I would be most grateful for your
> > help.
> > 
> > Cheers
> > Dianne Emslie
> > 
> > 
> > 
> 
> 
> 



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