Cloning problem

Brendan Scott brendans at
Mon Feb 3 20:43:51 EST 1997

We have been having an extremely dogged cloning problem.  We take microbial
genomic DNA, treated with CsCl, and do a partial digest with Sau3A.  We
ligate it into pUC 18 and transform it, and it works fine.  BUT we are
trying to make a library of fragments larger than 2kb, so we have tried
fractionating the partial digest with several different fractionation
methods, and then ligating, etc.  Regardless of fractionation method, we
can't get the cloning to work!

Does anyone have any suggestions?


Brendan Scott BSc. PGDip(Hons. (Parasitology)) Qld

Institute of Dental Research            Department of Pathology
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