Qiagen Maxi: Quantity

Zophonias O. Jonsson zjons at vetbio.unizh.ch
Fri Apr 19 08:21:33 EST 1996


In article <4l7kms$me0 at mailhost.zeneca.co.uk>, john brennand
<john.brennand at gbapr.zeneca.com> wrote:

> 
> However, it sounds like this isn't your problem - 200ug/ml is pretty 
> high

Just to put in my worthless IKr 0.02.  Couldn't this be the problem? 
200ug/ml can be quite a lot for a low copy number plasmid like pBR322 if
you are starting directly from glycerol stocks.  If I remember the
original post correctly the problem was with pBR322, not pUC.  I would try
to use less Amp, smaller culture volume and amplify the pBR322 with
Chloramphenicol.
Alternatively, could it be that you simply have a bad batch of columns? 
We did have a box that repeatedly gave us low yields but of course there
may have been other reasons for it.

Zophonias

_____________________________________________________________________
Zophonias O. Jonsson
Institut fur Veterinarbiochemie               Tel: (41-1)-257-54-75
Universitat Zurich-Irchel                     Fax: (41-1)-362-05-01
Winterthurerstrasse 190
CH-8057 Zurich
Switzerland
_____________________________________________________________________



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