Help! low recovery yeild of geneclean

mbxspd at unicorn.nott.ac.uk mbxspd at unicorn.nott.ac.uk
Sun Nov 20 16:52:24 EST 1994


Off the top of my head I seem to recall that:-
1) TBE gels stink for gene-clean (even with the addition of 1M
mannitol to aid gel solubilization we find TAE gels give better
yields).
2) A pH of 7.5 or lower seems to be good for the binding step. Too
high a pH yields a highly charged DNA and it won't bind well to the
glass.
3) Elution, conversely, needs a higher pH. We always elute in TE, pH8
at room temp. Usually works great.

A quick test to see where you're losing your DNA might be to gel
purify some 32P or 35S-labelled lambda/HindIII standards or something
using your kit.

Hope some of that is some help.


Simon Dawson
Dept. Biochemistry
Queens Medical Centre
Nottingham University
U.K.

Internet email: mbxspd at unicorn.nott.ac.uk



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