I'm looking for a method for getting good reliable plasmid preps
from Agrobacterium. In our lab we can get good plasmid preps from LBA4404,
A722, A4, and 9 other strains if they contain a pGA643-based plasmid or
pKIWI110. We can't get any reliable result if we try A281, EHA101, or C58,
or if we try LBA4404 or A722 containing a pBIN19 or pBI.121-based plasmid.
Speaking of pBIN19-based plasmids, how do you transform them into
Agro and then get the sodding things to grow? We find that a zapping in BRL'
s electroporator followed by 1-2 hours in YT or LB media before plating onto
selective media works fine for other plasmids but even mention the word "
BIN19" and things die.
Finally, in our hands we find that LBA4404 has a propensity to
insert a fragment of >2kb into pGA643-based plasmids from time to time. Has
anyone else ever encountered this problem?
Either E-mail me direct or better still post back here and start
some discussion going!
Plant Pigments Group
New Zealand Institute for Crop & Food Research
SMITHH at crop.cri.nz