No plasmid from Miniprep

[Zophonias O. Jonsson]

In article
<Pine.OSF.3.95.970103161057.30149A-100000 at saul3.u.washington.edu>, Nancy
Linford <nantzee at u.washington.edu> wrote:

> On 3 Jan 1997 KHUANG at CHEMVX.TAMU.EDU wrote:
> 
> =>Hi, Netters,
> =>I met a serious problem. I tried to isolate plasmid (pET series) from JM109
> =>by mini-prep, but I could not get any plasmids. The colonies were grown well
> =>on LB/amp(100 ug/ml). 

> The JM series are notoriously difficult to mini-prep due to a large number
> of cell wall carbohydrates. A similar cell line is the XL-1 blue and
> these are a much easier choice for plasmid mini preps.

> Nancy

Additionally the pET's are low copy number so one will always have rather
low yields.  I grow them in ultra rich, well buffered medium (Superbroth)
for minipreps.

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
_____________________________________________________________________