Southern Hybridization Buffers?
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
0.5M Na phosphate, pH 7.2 (0.5M in Na, so 134g/l of Na2HPO4+7H2O,
pH'ed with 4ml 85% phosphoric acid)
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.
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