rehybridization nylon membranes

Michael W. Thompson I.hate.spam at spam.hater.com
Wed Nov 19 21:10:24 EST 1997



Diederik van Tuinen wrote:

> What is the best method to wash a nylon membrane in order to make a other
> Southern Blot on it, and how often can you reuse a membrane (with
> radiolabelled probes)

I usually wash them in boiling 0.2x SSC, 1.0% SDS twice for fifteen minutes
(just pour the solution in with the blot and let it shake at room
temperature.  Then I check it with a geiger counter; if there's still a few
clicks, I do a third wash.  I reprobed one blot nine times this way; normally
they will be good for three or even four probings without a significant loss
in signal.

Michael Thompson
University of Kentucky
Department of Biochemistry

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