HELP:cloning into pGEM-T
bettenha at mpiz-koeln.mpg.de
Thu Aug 31 05:18:18 EST 1995
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,
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