Successful phagemid excision from Zap-express ???

Ravi Ramachandran rravi at u.washington.edu
Thu Mar 19 10:52:42 EST 1998


Dear Claire aka Smoothies: I have used the lZAP libraries with no
problem
(screened eight different libraries). Could there be a problem with your

helper phage?
Re. your second Q @ handling lambda DNA-the best way I hae found to
dissolve the lambda DNA in TE is when the pellet is still slightly wet.
Drying the pellet completely, always seems to make it more difficult to
dissolve.
Good luck with your excision,
Cheers,
Ravi

(No afffiliation with Stratagene or any of its subsidiaries)

Smoothies wrote:

> 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



--

************************************************************
Ravi K. Ramachandran
Senior Fellow
D-532, HSB, Box 357470
Department of Pathology
University of Washington
Seattle, WA 98195.

Phone : (206) 543-5363
Fax : (206) 543-3644
email : rravi@ u.washington.edu
Web: http://weber.u.washington.edu/~rravi/

************************************************************





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