Stripping blots

Sergio Torres setorres at nmsu.edu
Thu Dec 14 13:04:18 EST 1995


On 7 Dec 1995, Catherine Raven wrote:

> Hi, I have a protocol for stripping DIG labelled genomic southerns:
> Soln A:0.4NNaOH
> Soln B:0.1xssc/0.1%SDS, 200mM Tris-HCl pH7.5
> 
> 1) Incubate membrane for 30 mins in soln A, at 62 C .
> 2) Discard A and incubate in soln B, 30 mins at 62 C.
> 3) Wash filter in sterile 2x SSC and store sealed at 4 C.
> 
> I have used this for stripping DIG, and isotopically labelled blots and 
> been able to re-probe perfectly. I have stripped and re-probed upto 
> three times using this method, I have heard of people stripping more but 
> even Hybond N deteriorates eventually. Alternative is boiling in dH2O, 
> but this does wear the membrane significantly and you may lose your 
> bound DNA.
> Cheers 
> Catherine.
> 
  Catherine/Someone,
	Have you stripped RNA blots on positively charged nylon membranes
(Boehringer) & if so>> Did it work OKAY?.. Will you please let me know 
how you so it??? Thanks, Sergio...> > 



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