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Agrobacterium & Binaries

Howard R. Smith HRS at levin.cri.nz
Mon Jul 18 22:28:56 EST 1994


Hi netters,

	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.  
Any suggestions?
	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!

Howard Smith
Plant Pigments Group
New Zealand Institute for Crop & Food Research
Levin
New Zealand
SMITHH at crop.cri.nz



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