Northern hybridisation

David L. Haviland, Ph.D. dhavilan at IMM2.IMM.UTH.TMC.EDU
Fri Jun 18 12:39:27 EST 1999


At 09:48 6/18/99 -0700, Martin Dalziel wrote:
>Can anyone recommend an optimised method for northern hybridisation
>based on the Sodium Phosphate / 7% SDS technique?
>Specifically, membrane used, blocking etc..
>
Martin:

Yes, it is the method published in Biotechniques by Virca et al 8:370 (1990).
I use Hybond N+ but any good charged nylon will/should work.  I've been
able to reprobe upwards of 3-4 times, the transfer is light-years more
efficient that with the old 20XSSC protocol and I've obtained just a clean
a result as depicted inthe paper.  I never did or do a blocking step, I
simply hybridize with the probe in the same buffer used for the prehyb step.

Hope this helps,
David
=============================
 David L. Haviland, Ph.D.
 Asst. Prof. Immunology 
 University of Texas - Houston, H.S.C.
 Institute of Molecular Medicine  
 2121 W. Holcombe Blvd.  
 Houston, TX  77030 
 Internet:"dhavilan at imm2.imm.uth.tmc.edu" 
 Voice: 713.500.2413  FAX: 713.500.2424
 ------------------------------------------------------  
The hardness of butter is directly proportional to the softness
of the bread.
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