Best Hybridization Solutions for Southerns

Paul N Hengen pnh at ncifcrf.gov
Thu Apr 3 11:15:50 EST 1997


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:

@article{Hengen1996Octtibs,
author = "P. N. Hengen",
title = "Methods and reagents - Purification of {GST}
fusion proteins",
journal = "Trends in Biochemical Sciences",
volume = "21",
number = "10",
pages = "400-401",
month = "oct",
year = "1996"}

@article{Church1984,
author = "G. M. Church
     and W. Gilbert",
title = "Genomic sequencing",
journal = "Proc. Natl. Acad. Sci. USA",
volume = "81",
pages = "1991-1995",
comment = "church hybridization solution",
year = "1984"}

--
*******************************************************************************
* Paul N. Hengen, Ph.D.                           /--------------------------/*
* National Cancer Institute                       |Internet: pnh at ncifcrf.gov |*
* Laboratory of Mathematical Biology              |   Phone: (301) 846-5581  |*
* Frederick Cancer Research and Development Center|     FAX: (301) 846-5598  |*
* Frederick, Maryland 21702-1201 USA              /--------------------------/*
* - - -  Methods FAQ list -> ftp://ftp.ncifcrf.gov/pub/methods/FAQlist - - -  *
* -  TIBS column archive -> http://www-lmmb.ncifcrf.gov/~pnh/readme.html - -  *
* - The BEST Molecular Biology HomePage -> http://www-lmmb.ncifcrf.gov/~pnh/  *
*******************************************************************************



More information about the Methods mailing list