recipe for Church-Gilbert's solution

John Watson watson_j at bms.com
Thu Jul 24 16:28:37 EST 1997


Marc Rosoff wrote:
> 
> Hi everyone,
> I was wondering if someone could direct me to source for a recipe for
> this solution.  DOes it work much better than SSPE/DENHARDTS/SDS for
> Northerns?  I have heard that Nitrocellulose has much lower background
> for Northerns than Nylon.  Any truth to this rumour?

I don't know if this what you are loking for, but it's a modified
Church-Gilbert hyb-mix recipie (PNAS 81:1991-1995 (1984)). Gives low
background with nylon filters; works for Northers and Southerns. I'm not
convinced that it is the most *sensitive* hyb mix going.

                          For 200 ml:

BSA	              1%     (8 ml of 25%)
EDTA              1 mM   (0.4 ml of 0.5 M)
NaHPO4, pH 7.2    0.5 M  (50 ml of 2 M phosphate buffer)
SDS               5%     (40 ml of 25%)
Formamide         50%    (100 ml)
H2O                      (1.6 ml)

Cheers,

AJW
----------------
John Watson
Bristol-Myers Squibb Co.
watson_j at bms.com
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