DNA supercoiling

rogier stugerNOstSPAM at cellbiology.uni-frankfurt.de.invalid
Mon May 29 06:08:15 EST 2000


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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