chloroquine & trioxalen
stuger at cellbiology.uni-frankfurt.de
Tue Jan 25 08:48:22 EST 2000
we use chloroquine to see pBR322 topoisomers on gels.
We use 1.4 % agarose, and different chloroquine concentrations, ranging
from zero to 15 ug/ml in both gel and running buffer.
Our buffer is 1x TPE (10x TPE = 500 mM Tris, 550 mM H3PO4, 10 mM EDTA,
Make sure the chloroquine is well distributed in the gel: put a stirrer
in your agarose and spin it for a while.
Gels need to be run at low V and A (to keep the chloroquin alive).
For pBR322 on 14x11 cm gels we run 20 h at 50 V, 50 mA.
If your plasmid is smaller, you should run at lower power or shorter
time, otherwise your DNA will run out of the gel.
Mail me if you wanna know more.
Free U of A
rogier at bio.vu.nl
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