Qiagen Maxi: Quantity
Gerd.Romanowski at uni-bayreuth.de
Fri Apr 19 02:32:53 EST 1996
In article <4l52rh$jvb at mailhost.zeneca.co.uk>,
john brennand <john.brennand at gbapr.zeneca.com> wrote:
>Just a thought.
>Is it possible that the problem is in the cultures - ie the bugs dont
>contain as much plasmid as you think, because, for example, the
>antibiotic levels are too low or it has gone off and plasmid free
>segregants are overgrowing the culture ?
>You could check this by 1st doing a miniprep from the culture, by an
>alternative method, and then from the result predicting how much you
>should get from the whole culture.
I already did that: Qiagen minipreps are o.k. in quality and quantity.
Ampicillin is 200 µg/ml. That should be optimal. Is it a known "bug" with pUC
and pBR, that plasmidless segregants are overgrowing the culture? I dont't
think that this is a real problem because segregants without plasmid cannot
grow and the culture is logarithmic.
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