Southern Hybridization Buffers?

A. John Watson watson_j at bms.com
Thu Jul 13 13:36:06 EST 1995


In article <3tjhqj$jjd at mercury.hgmp.mrc.ac.uk>, veronica narvaez
<v-narvae at nimr.mrc.ac.uk> wrote:

> Hello Everybody!
> I'm going to start doing southerns, and I found there are a lot of different
> hybridization buffers, ie, some use a different prehybridization solution
> than the hybridization one. Some use Denhardt, some not, etc.
> Any preferred hybridiation solution? A nice, reliable, easy one?
> Any of you heard of "Church"  hybridization buffer? Is it good? Can 
> you give me the recipe?
> Thank you very much!
> Veronica

Here's mine, modified from Church & Gilbert PNAS 81,1991 (1984).

Reagent     [Final]     Stock
-------     -------     -----
BSA            1%         25%
EDTA           1 mM       0.5 M
NaHPO4, pH7.2  0.5 M      2 M
SDS            5%         25%
Formamide      50%        neat

IMO this mix reduces background at the cost of specific signal (that is, I
get stronger signals, albeit more background) with different mixes).

John

-- 
A. John Watson, Ph.D.                           | My opinions, of
Bristol-Myers Squibb Pharmaceutical Research    | course, are my own.
Central Nervous System Biology, Department 404  | Nobody else seems
Wallingford, CT 06492                           | to want them.
watson_j at bms.com                                |



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