Help

Michael Clarke mwclarke at uwovax.uwo.ca
Wed Jan 24 21:51:52 EST 1990


Hi!
Lately we've been having great troubles with our colony lift procedure.  We
are probing bacterial colonies for the expression of a cloned protein for
which we have monoclonal antibodies.  The problem is twofold:  i. with a
peroxidase detection system we see high backgrounds confined to all
colonies (even those transformed with plasmid with no insert).  ii. with an
alkaline phosphatase system we get no signal at all.  There's probably
more than one thing going on here, one may be that the colonies are not
lysing completely (we've been using 2% SDS at 70 C) another may be that
endogenous peroxidase is giving a signal, or ???  Could those of you who
have a successful colony lift protocol please send it along.  All
suggestions will be greatly appreciated.

Michael Clarke
Dept. of Microbiology & Immunology
Univ. of Western Ontario



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