Summary of responses to the request for methods for staining
tissue for GUS without killing the tissue: many thanks to all those who
unselfishly shared all their tips and/or references; you were a great help.
Three Plant Molecular Biology Reporter articles give methods for
non destructive GUS assays:
J.H. Gould and R.H. Smith (1989). A non-destructive assay for
GUS in the media of plant tissue cultures. Plant Molecular
Biology Reporter 7(3): 209-216.
Martin et al. Non-destructive assay system for detection of GUS
activity in higher plants. Plant Molecular Biology Reporter
G. Kirchner, C.J. Kinslow, G.C. Bloom and D.W. Taylor. (1993).
Nonlethal assay system of B-Glucuronidase activity in transgenic
tobacco roots. Plant Molecular Biology Reporter 11(4): 320-325.
In addition, Richard Jefferson comments in his Plant Molecular
Biology Reporter article (1987; vol5 no 4, p399) that
"reasonable staining can be achieved at much lower substrate
concentrations (50ug/ml) with enhanced recovery (of stained
Also, in a recent article, Swoboda et al. (1994) EMBO J. 13:
484-489 have an in vivo staining procedure they use on
Vacuum infiltrate for 5 minutes with sterile staining buffer
containing 200 mg X-Gluc in 300 ml 50 mM phosphate buffer (pH
7.0) 0.5 x MS medium. Incubate 4-8 hours at 37 degrees.
Martin et al. in Sean Gallager's book "Gus Protocols" - Academic
Press 1992 has a method for using MUG in intact tissues. We we
have used this technique to confirm that seedlings were
transgenic and it seems to work okay. We did not try to
continue growing the plants, but the authors suggest it is
possible (Prof. David Oliver , University of Idaho)
| Ed Rybicki, PhD | "Now you've got the hang of it |
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