A problem in topology

Ed Beaty ebeaty at firecraker.com
Fri Aug 9 20:41:44 EST 1996


I was wondering if anyone had a good way to completely convert a batch of
double-stranded, unnicked plasmid to single-stranded, unlinked closed circles?
I assume that if you heat a ds unnicked plasmid to denature it, the two 
ss circles remained linked, and quickly reanneal upon cooling. 
 
The catch is, I can't use a phagemid vector to directly make ss circular 
DNA in this case.  If anyone knows a good way to unlink two linked circles 
(using a topoisomerase, maybe?) I'd be extremely thankful to hear it.

thanks,
Edward Beaty
Pierce Chemical Company



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