pET vectors and low yield

mark damico mwdst1 at pitt.edu
Fri Jan 20 14:23:45 EST 1995


In article <D2J8uA.4yL at UQuebec.CA>, Benoit_Hebert at IAF.UQUEBEC.CA () wrote:

> Gilles VACHON (vachon at bio.grenet.fr) wrote:
> > 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.
> 
> As Cornelius pointed out, this is normal. Indeed, I get similar amounts with
> pMal-c2. I did get better results in XL-1 Blus when compared to TB1. I
haven`t 
> tried the DH5alpha yet, I will during the next week. I suggest to amplify your
> production with chloramphenicol.
> 
i have tried to amplify yields using chloramphenicol with pET-17B. It isn't as
great an increase as you might expect. I don't know why, exactly, but it
may be enough for your application. Just remember, you are smarter than
the bacteria.

-- 
mwd



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