Cosmid cloning problems

Volker Knoop volker.knoop at
Fri Sep 5 07:54:29 EST 1997

A question to people with cosmid cloning experience

We have tried to introduce ligations with cosmid pWE15 (Clontech) after
packaging with the Boehringer packaging extract into the ED8654 strain
supplied with the packaging kit.
We obtained absolutely no single clone.
Any obvious incompatibilities that we are not aware of?
Boehringer recommends washing the recipient cells with 10 mM MgCl2
before infection - while  Sambrook/Maniatis uses fresh grown cultures
(likewise grown in broth + MgCl2 and Maltose). Differences?

Any help is highly appreciated

Thanks, VK

PD Dr. Volker Knoop (mailto:volker.knoop at
Allgemeine Botanik, Universitaet Ulm, 89069 Ulm, Germany (Letters)
Address: M24, Albert-Einstein-Allee 11, 89081 Ulm
Tel +49-731-502-2615, Lab -2616, Fax -2626

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