[Microbiology] pBADabg

Brian Higgins higginsb at uga.edu
Fri Jun 9 14:32:56 EST 2006


Has anyone used Stewart's pBADalpha-beta-gamma plasmid in DH5alpha?  I'm
trying to use the galK selection method, so many of the other commonly used
E.coli strains are out b/c they already have a disrupted galK.  I'm having
real problems getting correct transformants (keep getting random mutants).
The only thing I haven't followed in their protocol is the electroporation
condition--I just used a common E. coli setting since out pulser won't allow
us to change many of the parameters.  the conditions i'm using work when
putting other plasmids into DH5alpha, so I figured they would work here.
Any suggestions?  I'm plating xformants onto 2-deoxygalactose minimal media
plates (after washing in M9 salts).
Thanks!


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