Northern hybridisation

Jil Tardiff jtardiff at westnet.com
Fri Jun 18 16:11:09 EST 1999


David,

My student has been struggling with Northerns of late, would you mind posting a
synopsis of the method you use? I have a decent Biotechniques library, but it
doesn't go back to '90!

Jil




"David L. Haviland, Ph.D." wrote:

> 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.
>




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