Stripping blots

Catherine Raven craven at molbiol.ox.ac.uk
Thu Dec 7 13:15:06 EST 1995


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.





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