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Arbidopsis Callus

Fejes ErzsThetabet Fejes.nb.szbk at nucleus.szbk.u-szeged.hu
Thu Jun 1 00:37:27 EST 2000


Hi Mauro!
     This is how I did it:
     Prepare Callus Inducing Medium (CIM) in Petri dishes:
         MS salts
         2% sucrose
         pH adjusted to 5.7-5.8 with KOH
         0.2 mg/l kinetin
         1 mg/l 2,4-D
         0.2% Phytagel
     Prepare AM medium, also in Petri dishes:
         0.5x MS salts
         1% sucrose
         pH to 5.7-5.8 with KOH
         0.2% Phytagel

     Germinate sterilized Arabidopsis seed on AM medium, in the light.
After 10 days you should have seedlings with expanded cotyledons and
the first pair of true leaves just starting to show. Excise leaf
and/or hypocotyl pieces (you should do this under water in a Petri
dish, otherwise the seedlings dry out before you are finished) and
lay them on solidified AM medium. Place them in light and callus
growth should start soon.
     If you need more details, please ask - I cannot know what your
previous experience is with this type of work.
     Good luck:
     Zso Fejes
     Institute of Plant Biology
     Biological Research Center
     H-6701 Szeged, P.O.Box 521.
     Ph. 36-62-432080/287
P.S. Your e-mail address doesn't seem to accept messages sent using
the reply function.


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