Re: Problem in selection, please suggest.

[First Name Last Name <juamber@MY-DEJA.COM>]

Assuming that transformation is taking place and that the target tissue, selection regime 
and medium used are OK, looks like an excess of bacteria on the tissue during/after the 
cocultivation phase. I would propose to:
check/reduce the amount of bacteria used for infections,
check/reduce infection and/or cocultivation times,
check/improve the washing step to remove bacteria from the tissue after cocultivation,
check/change/adjust the antibiotic used for killing your agrobacterium strain.
Success,
Juan B.

>Date:         Thu, 8 Feb 2001 00:43:14 -0800
>Reply-To: Plant Tissue Culture <plant-tc@tc.umn.edu>
>From: Boosani Chandra shekhar <boosani2001@YAHOO.COM>
>Subject: Problem in selection, please suggest.
>To: plant-tc@tc.umn.edu
>
>Dear all,
>I am working on trasformation of indica rice using
>Agrobacterium.
>I am transfering the cocultivated callus to selection
>medium containing Cefotaxim at 500mg/L and Hygromycin
>at 50mg/L along with 3mg/L concentration of 2,4-D.
>All the time i could find the callus turning into pale
>brown and are not proliferation even after 6 weeks on
>selection.
>please suggest with some clues that may help me in
>proliferation the transformed callus on selection
>medium.
>
>Thanking you in advance
>Chandra Shekhar

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