Ligation problems

Stephen Gately sg at nwu.edu
Thu Mar 16 07:30:27 EST 1995


In article <D5FG0n.1qM at demon.co.uk>, Anthony Tomlinson
<tomlinson at pplros.demon.co.uk> wrote:

> In article <3js36s$e27 at thot.u-strasbg.fr> Francois Nantel,
> Nantel at titus.u-strasbg.fr writes:
> >Dear bionetters,
> >
> >I presently have big ligation problems but that was expected. I need to
> >put a 3kb insert into a 3.2 kb vector. Oh, did I tell you it is a blunt
> >ligation :-0
> >
> >I tried treating the vector with alkaline phosphatase but the ligation
> >effeciency is so low I didn't get anything.
> >
> >If anybody have a knock-out technique for hard to do ligation, I would be
> >very grateful.

Have you tried Library Efficiency DH5 alpha competent bacteria (GIBCO
BRL).  These improved the number of white colonies I obtained when
ligating a blunt insert into a CIAPed vector.

Good luck



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