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isolating large plasmids - is it possible (Aug/26/2006 )

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thanks for your replies.
what do you think if PFGE( Pulse Field Gel electrophoresis) is done? which will be better- PFGE, CsCl2 or Bacmid protocol .
which will more easy and less time consuming?


You probably have to use PFGE in order to resolve plasmid of this size. I think most purification protocols may not be feasible for it, because giant sized DNA are very easy to break down even by pipeting.


well, as the bacmid prep is not hard to do, i would try it. But for checking your prep, only PFGE will resolve it. I think it's less damaging than PFGE and DNA extraction from the gel.
Electroelution from the gel is the only method here applicable, and for precipitation, butanol should be used as the mix water + alcohol can be done by inverting the tube and spin is done 2' at 1000 rpm. it's not that dammaging.


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