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.



More information about the Microbio mailing list