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.
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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