In article <belville-2410961404200001 at cherubin.ens.fr>,
belville at wotan.ens.fr (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°C).I put 5 or 20 more inserts than
> plasmid.
> Is it necessary to change of vector? is it better to work at +4°C?
>> Thank you for your answers
>>> Corinne Belville
>> --
> Daniele EUSEBE(Ecole Normale Supérieure-INSERM U293-Paris-France)
> e-mail:deusebe at wotan.ens.fr
Salut Corinne,
most important thing is: have you used Taq polymerase? If not, using pGEM-T
will never work. What about your control without inserts? Have you tried
CIPing the vector?
Marc
