pET vectors and low yield

sb9e sb9e at faraday.clas.virginia.edu
Mon Jan 16 12:32:27 EST 1995


In article <3fdfm9$hhn at mserv1.dl.ac.uk>
vachon at bio.grenet.fr (Gilles VACHON) writes:

> Dear netters,
> 
> I am having trouble in getting regular amounts of pET (19b or 11a, with ot
> without insert)) in DH5alpha.
> I have only 30-50ug of plasmid from a 200ml LB culture  (Cesium Chloride prep).
> 
> The DH5alpha strain is ok since I get 500-700ug of pUC in the same conditions.
> 
> Does anyone on the net have an answer to that problem ?
> I am planning to do cloning with these pETs and I bet I won't be able to
> check the clonings with that low yield.
> 
> Thanks
> Dr. Gilles Vachon
> Laboratoire de Biologie Moleculaire Vegetale


Same with me. pET is a low copy number plasmid. Did anybody try
chloramphenicol amplification? 

Serge



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