root-specific protein/RNA method

Jan Traas traas at TOURNESOL.VERSAILLES.INRA.FR
Fri Nov 19 09:52:54 EST 1993


We have been using methacrylate embedded seedlings of Arabidopsis for both in
situ and immunofluorescence studies. The immunofluorescence  method is
described in detail by Baskin et al. (Planta 187 (3) 1992) and we have
slightly modified the procedure for in situ analysis (Kronenberger et al.
Cell Biol. Int. in press). The method is ideal for Arabidopsis. If you
need further information on the procedure, please let me know. At least
in our hands most other procedures have given poor tissue preservation.
Whole mount in situ hybridization works fine for young parts of the root,
but we have had problems with the more mature tissues which are impossible
to label. Moreover, tissue preservation is poor. For immunolocalization we
also use plantlets that have been split in two longitudinally after
fixation and cellulase treatment. These preparations can only be studied
using confocal microscopy.

Good luck!

Jan Traas





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