DNA supercoiling

Dr. R O'Kennedy r.o'kennedy at ucl.ac.uk
Thu Jun 1 06:25:31 EST 2000

Rogier and Susanne,
Thanks for the advice. The other thing is I presume that standard 
activated carbon is enough to decontaminate chloroquin containing 
solutions prior to disposal.


rogier wrote:
> Hi Ronan,
> for pBR322 I use 1 to 15 ug/ml chloroquin in gel and running
> buffer.
> Topoisomers of pBR isolated from E. coli are clearly separated at
> 10 ug/ml chloroquin, except when the DNA is extremely relaxed
> (like when you add a gyrase inhibitor), then you need to add less
> chloroquin. Extremely supercoiled DNA needs more chloroquin to
> pull the bands apart. but not too much, or you'll get positively
> writhed plasmids on top of the negative ones.
> Good luck,
> Rogier
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Dr. R O'Kennedy
Senior Research Fellow
The Advanced Centre for Biochemical Engineering,
Department of  Biochemical Engineering,
University College London,
Torrington Place,
London WC1E 7JE
e-mail r.o'kennNOedySPAM at ucl.ac.uk
phn 020 7679 3247

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