Plasmid isolation
Jay Mone'
jmone at MARAUDER.MILLERSV.EDU
Fri Mar 27 09:16:49 EST 1998
Microbabe,
I get pretty good yields of plasmid from B. sub using the alkaline
lysis method described by Maniottis in Molecular cloning. The only
modification I use is that I add lysozyme after I resuspend the
bacteria in solution I, and incubate at 37oC for 10 minutes before
adding solution II. The amounts I get are about 20% of the usual
yield for a plasmid like bluescript in DH5alpha cells, but that might
be related to copy numbers, rather than technique.
But while I have your attention, my problem is in the transformation
protocol. My efficiencies are very poor. How do you transform your
B. sub?
Jay M.
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