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stripping Southern blots

Pamela Lagali plagali at gpu2.srv.ualberta.ca
Thu Oct 17 20:53:55 EST 1996


I am looking for a protocol for stripping (nylon) Southern blots.  I have tried
using a boiling 0.1% SDS solution poured over the blot and allowed to cool to
room temperature (i.e. the "harsh treatment" for stripping as prescribed by
"Current Protocols in Molecular Biology").  This method removes most of the
hybridized probe, but some of it remains in addition to a significant amount of
background signal.  I performed the above procedure twice, and there was no
appreciable reduction in signal intensity when the blot was re-exposed to film
for the second time.  I am confident that the blot was not allowed to dry out at
any point (but I can never be sure I guess).  Does anyone know of any HARSHER
stripping procedures which I could try to attempt to remove what remains of the
hybridized probe? 

Thanks for any suggestions,


Pamela Lagali
Department of Cell Biology
6-50 Medical Sciences Building
University of Alberta
Edmonton, Alberta, Canada
T6G 0X6
ph: (403) 492-7407

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