Castor Menendez (menendez at mibm.ruf.uni-freiburg.de) wrote:
: Dear netters,
: Who can recommend me a reliable method for preparative scale
: purification of a large plasmid ?
: I am working with a 160 KB plasmid from a Gram + organism.
: We have tried a variety of methods with very disappointing results.
: All are labor intensive and give very poor yields
: (5 ug / lt culture approx.), or no plasmid at all.
: Does anybody have tried something like large scale plasmid
: purification using pulsed field electrophoresis?
: Thanks for your help.
: C. Menendez
: (menendez at sun2.ruf.uni-freiburg.de)
I suppose you have the Kado and Liu proceedure. The proceedure has some
technical tricks to it, but was used to isolate the Ti plasmid of
Agrobacterium tumefaciens (200 kb). I have never tried it on a gram +
bacteria. Contact me for further details re Kado and Liu.
Mike Cooley mbcooley at ucdavis.ucdavis.edu