[Arabidopsis] BASTA concentration of JatY clones
Jose M. Alonso
jmalonso at pop-in.ncsu.edu
Thu Mar 30 14:40:08 EST 2006
We have been selecting JAtY transformants in MS and 1% sucrose plates
supplemented with 50mg/ml of phosphinothricin (glufosinate ammonium from
Grescent Chemical Co.). For an easier/faster identification of the
transformants, we stratify the seeds at 4C for three days in the plates, and
then let them germinate for three days in the dark at 22C. Plates are then
moved to constant light: transgenics can easily be identified after 24h.
Jose M. Alonso Ph.D.
Department of Genetics
North Carolina State University
Raleigh, NC 27695
Phone: (919)515 5729
Fax: (919) 515 3355
Email: jmalonso at unity.ncsu.edu
I have recently transformed my plants with JatY library clones (obtained
from GetCID) which were constructed using the pYLTAC17 vector, allowing
for the selection of transformed plants by BASTA resistance.
Ideally I would like to select for my transformants on soil by spraying
them with BASTA. However, I am struggling to optimize the BASTA
concentration required for selection. Could anyone who has had success
with these clones please tell me what concentration of BASTA/glufosinate
ammonium (or phosphinothricin for selection on plates) to use?
Secondly, if someone has plants successfully transformed with any of the
JatY clones that I could use as a control in my selection process, I
would be most grateful if you are willing to send some seed to me.
Please see the postal address below.
Dr Katherine Denby's lab, room 430
Department of Molecular and Cell Biology
University of Cape Town
Tel: +27-21-650 4006
Fax: +27-21-689 7573
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