quick hybridizations

Estelle Hrabak HRABAKE at MACC.WISC.EDU
Wed Mar 2 22:59:10 EST 1994


>Hi, Does anyone know of a recipe for hybidization cocktail that is quicker
>than the standard formamide - dextran mixture?  I've been using
>Formamide/20xssc/2M tris/100x denharts/water/10% sds/50% dextran
>(92/48/1/2/11/18/40) and it's pretty slow.
>
>Thanks   Stephen Morris
>smorris at pharma1.pharma.mcgill.ca
>
I use 50% formamide/6-7% SDS/0.25 M NaPO4 buffer (from a 1 M stock of NaH2PO4
pHed with phosphoric acid to pH 7.2)/1 mM EDTA at 42 C.  Wash with 0.25 M
NaPO4/1-2 % SDS/1 mM EDTA at 42 or 65 depending on desired stringency.  42 C is
usually fine for me.
P.S.  I hope I remembered the right sodium phosphate above-I don't have the
recipe here.  It's the dibasic.
Estelle



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