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megaplasmid megaprep

szcooley at chip.ucdavis.edu szcooley at chip.ucdavis.edu
Mon Jul 18 10:39:30 EST 1994


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




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