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!
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).
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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