Southern Hybridization Buffers?

Tjian lab insomnia at mendel.berkeley.edu
Sun Jul 9 19:45:01 EST 1995


Here's my recipe for Church's buffer.  IMHO, it's the best general 
purpose hyb buffer I've ever tried.  Low background, easy to use, nice 
signals.

0.5M Na phosphate, pH 7.2 (0.5M in Na, so 134g/l of Na2HPO4+7H2O,
                            pH'ed with 4ml 85% phosphoric acid)
7% SDS
0.5mM EDTA

No salmon sperm or herring sperm DNA needed, no Denhardt's, use for both 
pre-hyb and hyb.  Because of the 7% SDS, though, the DNA should be 
crosslinked to the membrane with UV.  Drying on Amersham's hybond-N+, 
however, appears to be sufficient.

Shane Albright
MCB Dept
UC Berkeley





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