Successful phagemid excision from Zap-express ???

Smoothies channels at sghms.ac.uk
Thu Mar 19 08:12:37 EST 1998


Hi

I have just tried to perform single clone excision of the pBK plasmid
out of a lambda (Zap-express) clone
and although I managed to excise something, it had nothing to do with
the original lambda clone...
Has anybody out there done excision routinely and successfully ?
I know I am not the only one having problems with excision, I'd like to
know what's happening. Is there some kind of recombination ? Does it
depend on the host cell ? (I am using the recommended XL-1 and XLOLR)
Stratagene are not being helpful at all and in fact they seem to keep
very quiet about it - is there a flaw in the
supposedly "simple and efficient" procedure ?
ANY help would be greatly appreciated as I am banging my head against
the walls by now.

I would also like to know if there are any tricks to dissolve lambda DNA
in TE ? Excuse my ignorance but
I am new to working with lambda.

Many thanks

Claire Fenech - PhD Student
Department of Pharmacology
St George's Hospital Medical School
London - UK
Email: c.fenech at sghms.ac.uk




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