Stripping Northerns

Klaus Salger salger at wap4.zi.biologie.uni-muenchen.de
Mon Feb 10 06:15:11 EST 1997


Martin Cann (mjcann at MAIL.MED.CORNELL.EDU) wrote:
: I have a Northern which has a high background.  The nylon has not dried
: out at any point and I have tried boiling 10mins in 0.5% SDS to remove
: the background but to no avail.  Does anybody know how to strip this
: Northern or does it just have to sit in the freezer for a month?

: Martin Cann

: mjcann at med.cornell.edu


Martin,

I found my formamide containing washing buffer to be more efficient for
stripping northerns than boiling the blot. Incubate the blot at 70 degrees C
for 2x 30min. (I do the stripping just like a hybridization in hybridization
tubes in a hybridization oven).
I guess any buffer with 50% formamide will work. If you want to try mine
here's the recipe:
        50%     formamide
        2x      SSC
        1%      Sarcosyl
        2mM     EDTA
        0.2%    SDS
        120mM   phosphate buffer pH6.8

If this doesn't work I would actually put the blot in the freezer.


Cheers
  Klaus

--
Klaus Salger		phone : +49 (0)89 5902 	-502
Zoologisches Institut	FAX   :			-450
AG Wetterauer		e-mail: salger at zi.biologie.uni-muenchen.de
Luisenstr. 14
80333 Muenchen
Germany

BioLinks: http://www.zi.biologie.uni-muenchen.de/~salger/salger.html



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