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).