cloning problem...

Dr. Roland Barten rb248 at mole.bio.cam.ac.uk
Mon Jul 31 04:00:56 EST 2000


We have been trying to clone a human gene in to TA vector. We've got
blue/white selection,  have screened the white ones directly by PCR, and
then tried to grow up those with inserts - this is where the problem
starts. On trying to grow up for mini-preps, nothing happens. There is
absolutley no growth.
We have tried different media and both Kan and Amp selections (as well
as no antibiotics at all), but we just cannot get any of these colonies
to grow in liquid media - except for once when it turned out that the
insert had two base pair changes.
One option would be phage - any other suggestions?

Thanks
Anne-Marie Lucas
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