Northern blot Pb

ORAC ORACII at aol.com
Thu Oct 15 09:49:01 EST 1998


In article <199810142046.OAA180436 at nestor.NMSU.Edu>, hroychow at NMSU.EDU
(Hiranya Roychowdhury) wrote:

>        A lot of things may go wrong. Your post does not elborate on the
>conditions of hybridization etc. (ie. the soln. used, temp., probe conc.).
>Room temp wash may or may not be stringent depending on the ionic strength
>of the wash. Also, whether you get signals on the blot depends on the
>presentation of the target messages. Why not run a positive control on the
>gel to be sure that that everything else is OK. 
>        Anyway, after the recent positive feedback about Church & Gilbert's
>protocol, I tried it for both Southern and northern simultaneously (Hybond
>NX). Seeing the results, I must confess that I will try not to revert back
>to the system using Denhardt's / formamide etc.

I, too, use the Church and Gilbert protocol. It's far superior to
Denhardt's solution, although the buffer is a bit of a pain to make (all
that SDS, you know).

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