Path: utzoo!utgpu!watserv1!watmath!uunet!bionet!lars.afrc.ac.uk!PHILLIPSA
From: PHILLIPSA@lars.afrc.ac.uk
Newsgroups: bionet.molbio.methds-reagnts
Subject: (none)
Message-ID: <9008101013.AA27286@genbank.bio.net>
Date: 10 Aug 90 09:14:55 GMT
Sender: daemon@genbank.BIO.NET
Lines: 18


Does anyone out there have experience with lambda ZapII? I'm
having great trouble getting the in vivo excision protocol to
work. I am following the method described by Stratagene exactly,
and I've titred my lambda and helper 'phage stocks to check that
they are within the required range. I've tried a 100,000 fold
range in the ratio of helper to lambda, and I've tried incubating
the Amp selection plates at 42degC to prevent co-infection with
helper. Still no colonies (well perhaps one or two occasionally).
It's nothing to do with the inserts, I get the same result with
straight lambda Zap2.
Any suggestions?

Andy Phillips
AFRC-IACR Long Ashton Research Station, Bristol BS18 9AF, U.K.
EARN/BITNET: PHILLIPSA @ LARS.AFRC.AC.UK
TEL: (+272) 392181 ext 254
FAX: (+272) 394007