Culture of E.Coli (SURE) / Large Plasmids / Foreign Sequences
rufer at novsrv1.pio1.uni-heidelberg.de
Tue Apr 2 11:19:55 EST 1996
In article <4jq6b7$hfd at news1.h1.usa.pipeline.com>,2 Apr 1996 03:19:03
GMT,mkoch at usa.pipeline.co wildly stated/shouted/said...
>On Mar 30, 1996 07:48:31 in article <Culture of E.Coli (SURE) / Large
>Plasmids / Foreign Sequences>, 'ucklw08 at ucl.ac.uk (Dr Alexander J Annala)'
>>Sambrook/Frisch/Maniatis Molecular Cloning page 1.21 suggests that
>>where plasmids grow poorly because of their large size or because of
>>foreign sequences they carry it may be worthwhile to consider some
>>alternative ways to grow and treat a bacterial culture.
>>I have a BlueScript SK- derived plasmid containing a 12 KB mammalian
>>intron which yields very small amounts of miniprep DNA (from 2 ml TB
>>overnight cultures 50 ug/ml ampicillan). The culture also appears
>>to be unstable in the sense that nothing grows in a new culture tube
>>(2 ml TB with 50 ug/ml ampicillan) innoculated with a sample drawn
>>from the previous culture. It would seem the cells no longer harbor
>>What alternative ways to grow and treat the bacterial culture should
>>be considered in the instant situation?
>>Thanks, AJ Annala
may i humbly suggest our publication in biotechniques entitled:
"improved expression of toxic proteins in e.coli" ?
the suggested "plating" method may very well solve your problems,
as it concerns itself with extremely instable plasmids.
biotechniques, vol. 19, number 2, 1995, Suter-Crazzolara and Unsicker
More information about the Methods