bundock at rulsfb.leidenuniv.nl
Thu Oct 12 19:00:39 EST 1995
<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
>that I do?
>Firstly, it is very inconsistant in growth, sometimes growing
>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
>of comparable size. All conditions and cells identical.
>I use the strain Agli and have not tried others. Is there a difference
>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.
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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