PBR322 - maximizing yields

Dr R. Dalgleish ray at uk.ac.le
Thu Aug 27 10:47:29 EST 1992

> What are the best tricks for increasing yields of pBR-based
> vectors?
> Brian
The way which I used to amplify pBR-type vectors was this:

Add chloramphenicol to a final concentration of 200 micrograms per ml to
late log phase cells. If you add it too late it won't help. For large size
cultures you can weigh out the powder and just tip it in and shake. Folklore
says not to shake too vigourously. If you are working with smaller culture
then make a 34 mg/ml solution of choramphenicol in 50% ethanol and add the
appropriate amount.

Of course, be sure that your vector does not carry a Cm resistance gene! If
it does then you can amplify using spectinomycin. The amount to use escapes
me at the moment.

Raymond Dalgleish.

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