Best Hybridization Solutions for Southerns

Alexander Kraev kraev at bc.biol.ethz.ch
Fri Apr 4 09:01:54 EST 1997


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.

Cheers,

-- 
Alexander Kraev, PhD
Biochemie III, ETHZ Zurich
Phone 41-1-632-31-47
Fax 41-1-632-12-13
e-mail kraev at bc.biol.ethz.ch



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