low copy plasmid isolation

Jay T. Myers jtmyers at umich.edu
Wed Mar 1 09:45:38 EST 2000


Here's a protocol I've used in the past.

1. Grow bact. to an O.D. 590 of 0.8 - 0.9

2. Add powdered Chloramphenicol to a final concentration of 170 ug / ml and
incubate again o/n at 37o C.

3.  Continue with whatever plasmid isolation you prefer.

Note:  The Chloramphenicol stops the bact. from dividing but still allows
for DNA replication therefore amplifying the plasmid of interest.  Hope this
helps.

Jay
> From: neweb at alcor.concordia.ca ( ELAINE B S NEWMAN )
> Organization: Concordia University, Montreal, Canada
> Newsgroups: bionet.molbio.methds-reagnts
> Date: 29 Feb 2000 20:26:20 GMT
> Subject: low copy plasmid isolation
> 
> does anyone have a method for isolating enough plasmid from a
> low copy vector to determine its structure by restriction enzyme cutting?
> 
> EBNewman





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