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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