HELP:cloning into pGEM-T

Berthold Bettenhausen bettenha at mpiz-koeln.mpg.de
Thu Aug 31 05:18:18 EST 1995


Dear Netter´s,
Over the last weeks I´m trying to clone a 1.6 kb PCR-fragment into the 
pGEM-T vector from Promega. I didn´t get any positive clone. In nearly
half of the minipreps the vector is even smaller than 2.4 kb. 
I think I have checked all which could go wrong. I ordered a new kit, 
made new competent cells, tried to ligate from several PCR-reactions,
eluted the fragment before ligation. The ligase is doing well in other 
ligations, so that might not be the problem.
I have to clone that particular fragment, because I can not use primers
with restriction sites.
Is there anybody out there who can help me???

Thanks for your advices,

Berthold Bettenhausen



More information about the Methods mailing list