Best Hybridization Solutions for Southerns

Robert Saunders rdcsaunders at dundee.ac.uk
Fri Apr 4 19:02:54 EST 1997


Alexander Kraev wrote:
> 
> In article <5i0l3m$12118 at ncisun1-nf0.ncifcrf.gov>, pnh at ncifcrf.gov (Paul N
> Hengen) wrote:
> 
> > paul gold (paulgold at MO.NET) wrote:
> >
> > > Our lab is gearing up to do Southerns in order to analyze genomic DNA
> > > from potential homologous recombinant ES cells for a knockout.  There
> > > are a lot of mixes, both commercial and in manuals, that claim maximum
> > > sensitivity with low background.  We would greatly appreciate feedback
> > > from those of you that have a favorite hybridization solution.  Thanks.
> >
> > If you plan on using the positively charged nylon membranes such as
> > Nytran Plus, Hybond N+, Magnacharge, BioDyne A, etc., you are probably
> > better off using Church's formula for the hybridization. The people on
> > this group say that it reduces the background considerably, especially
> > with the new Hybond N membranes that have changed formulation recently.
> > If you use a different hybridization solution, avoid adding formamide.
> > See the section on "Churchgoers" in this article:
> <snip>
> I'll second this, being a long time Churchgoer. I also use this solution,
> with 1% Boehringer blocking agent in place of BSA, for DIG/biotin
> hybridizations.

For 32-P labelled probing, I don't use BSA in Church, and it workes
fine.

Robert



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