Stripping blots

Antoine VEKRIS vekris at u-bordeaux2.fr
Wed Dec 6 03:54:07 EST 1995


In article <c.okane-0512950953280001 at cok-mac2.gen.cam.ac.uk>,
c.okane at gen.cam.ac.uk (cok at mole.bio.cam.ac.uk) wrote:

: Hi,
:    We have a few valuable blots, mainly hybond-N, with DNA bound. What's
: the best way to strip off a probe with minimum wear and tear to the blot
: and the immobilised DNA? We use DIG-labelled probes, so the stripping has
: to work well too, because we can't just wait for the probe to decay.

Hello there,
I have re-use southern blots for up to 41 times, using DIG-probes,
chemiluminescent detection and stripping off the probe by boiling for 2
min in a 2% SDS solution in water. A lot of foam but a great result. There
is a very important point however conserning the quality and amount of
probe necessary for hybridisation. Dont overload your system, filter the
probe solution throught a .45 micrometer filter and check for absence of
background by incubation of a small membrane  and revelation. If
background occurs just dilute a little bit more your probe.

Good luck Cahir

: 
: Thanks,
: Cahir
: cok at mole.bio.cam.ac.uk
: 
: -- 
: Cahir O'Kane, Dept of Genetics, Univ. of Cambridge, UK
: cok at mole.bio.cam.ac.uk

-- 
Antoine VEKRIS
Molecular Pathology and Gene Therapy group  /  University of Bordeaux II




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