Plasmid isolation

Fri Mar 27 09:16:49 EST 1998

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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