Ligation problems

Emmanuel Skoufos, Ph.D. skoufos at nefeli.imbb.forth.gr
Sat Mar 11 17:22:38 EST 1995


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.


Try using the blunt end ligation buffer discribed in the Maniatis text.
It has been working really well for me on stubborn ligations :-)

Cheers,
Emmanuel



_______________________________________________________________________________

Emmanuel Skoufos, Ph.D.
Insect Molecular Biology Group
Institute of Molecular Biology and Biotechnology
Box 1527 
Heraklion, Crete 71110
Greece



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