Ligation problems on agarose-isolated fragments

Dr. Arthur L. Kruckeberg bmbckh at
Fri Feb 2 08:42:47 EST 1996

I had a similar problem with ligation. I digested DNA separated on
multi-purpose grade agarose and cut out the required fragment. DNA was
extracted using Qiagen gel extraction kit. DNA was recovered as seen by
agarose gel electrophoresis but the DNA could not be ligated into our
vector. The agarose used was changed to ultrapure grade and isolation
of DNA was repeated. Ligation was successful. The source of your
problem maybe the agarose as it was in our case.
All the best



Chris Hoyle

   Department of Biochemistry and Molecular Biology
University of Leeds             phone +44 0113 2333172
Leeds LS2 9JT                   FAX   +44 0113 2333167
Great Britain           e-mail bmbckh at

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