Ligation problems

Thomas Burmeister tbu at gmx.net
Sat Aug 8 13:53:36 EST 1998


In article <6pl4mh$bb1 at panix3.panix.com>, iayork at panix.com (Ian A. York) wrote:

>We're having a terrible time getting what should be a straightforward
>ligation to work.  I have a feeling we neglecting something obvious, so I
>won't be offended at simple suggestions ...
>etc. etc...........


Hello Ian !

Just an idea: you may try to use a different bacterial strain for
transformation. I can remember a similar problem, that I experienced some
years ago. At that time it seemed to be just impossible for me to get a
1.5 kB insert into a bacterial vector ("pBacPak9", Clontech) designed for
constructing recombinant baculoviruses. I got plasmids with inserts, but
with the wrong size.
When I decided to change the bacterial strain (I had used E. coli XL
supercompetent bacteria from Stratagene) and to use E. coli DH5 alpha
competent cells, everything suddenly worked fine.

Good luck!

Thomas Burmeister
Free University Berlin




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