Agrobacterium problems

Paul Bundock bundock at rulsfb.leidenuniv.nl
Thu Oct 12 19:00:39 EST 1995


In article 
<Pine.OSF.3.90.951011100618.5908A-100000 at central.murdoch.edu.au>, 
wylie at CENTRAL.MURDOCH.EDU.AU says...
>
>Does anyone else working with Agrobacterium tumefaciens have the 
problems 
>that I do?
>Firstly, it is very inconsistant in growth, sometimes growing 
vigorously 
>in liqiud medium, and at other times growing slowly.
>Transforming it with large plasmids by electroporation or triparental 
>mating mating has been easy with some plasmids and impossible with 
others 
>of comparable size. All conditions and cells identical.
>I use the strain Agli and have not tried others. Is there a difference 
in 
>ease of use between strains? Where can I buy other strains?
>I store a glycerol stock at -80 C and plate onto LB + glucose every 
>couple of weeks, working from single colonies taken from that plate. 
>Any suggestions most welcome. Thanks.
>Steve Wylie
>Murdoch University
>Western Australia
>
I'm not sure how much trouble you are having with Agrobacterium in 
liquid culture but one antibiotic which can be a problem is gentamycin, 
especially in minimal medium.We mainly use strains with the C58 
background and these grow pretty much OK o/n in LB/LC without glucose.A 
nice review of Agrobacterium electroporation just came out:
"Electroporation of Agrobacterium", Methods in Molecular Biology,Vol 55,
  :Plant Cell Electroporation and Electrofusion Protocols".Humana Press 
Inc. Hope this is useful, Paul Bundock.




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