stripping Southern blots

[Michael Cooley]

Pamela Lagali (plagali at gpu2.srv.ualberta.ca) wrote:
: Hello,
: 
: 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,
: 
: Pam
: 
: 
: 
: Pamela Lagali
: Department of Cell Biology
: 6-50 Medical Sciences Building
: University of Alberta
: Edmonton, Alberta, Canada
: T6G 0X6
: ph: (403) 492-7407
: 

I'm convinced that harsher conditions are ineffective. Please see my post
to you via email and my post on this bullitin board. Thanks, Mike Cooley