Cloning problem

Brendan Scott brendans at med.su.oz.au
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?

Thanks,
Brendan.

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

Institute of Dental Research            Department of Pathology
7th Floor                               5th Floor
United Dental Hospital                  Blackburn Building
cnr Elizabeth & Chalmers Sts            University of Sydney
Surry Hills NSW 2010                    Sydney NSW 2006
Australia
(02) 9282 0352                         (02) 9351 3319
Fax (02) 9282 0368                      Fax (02) 9351 3429
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