On Thu, 24 Oct 1996, Corinne BELVILLE wrote:
> I would like to realize a ligation of a fragment whom the size is 8kb. My
> ligation failed in vector pGEM-T (at 15=B0C).I put 5 or 20 more inserts t=
> Is it necessary to change of vector? is it better to work at +4=B0C?
> Thank you for your answers
> Corinne Belville
> Daniele EUSEBE(Ecole Normale Sup=E9rieure-INSERM U293-Paris-France)
> e-mail:deusebe at wotan.ens.fr>=20
I have ligated similar large fragment to vector,my condition is 15=B0c,10=
ul volume,T4 1ul(5u),PEG8000 1ul, for 20 hours. It is blunt end ligation=20
. The key, I think, is to treat the fragment and vector properly.
Yu Yan Xing Ph.D
Unite Virologie and Imminologie Moliculares>of INRA