Ligation problems

Anthony Tomlinson tomlinson at pplros.demon.co.uk
Tue Mar 14 06:02:47 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.

Last time I had a hard blunt to do, I tried Ready to Go dried down ligase
from
Pharmacia, and it worked a treat.  It's not cheap, but they sent me a
free sample which was enough for
my experiment!

Anthony



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