Screening of Arabidopsis by Fluorescene assay

[REGCHAK at USTHK.UST.HK]

Dear netters,

     Does anybody read the paper from Bowling S A et al 1994, it showed the
fast screening of the overexpression of GUS activity by the fluorescence
assay.  The method is as follows

Incubation a single leave of 5-day-old seedlings in a microtiter plate
containing 100uL MUG at 37 C. After 24 hours of the incubation, the
fluoresacene of the overexpressor mutant were examinated under the long UV
lamp.

   
     I try to work on screening the GUS overexpressor by using this
protocol, however no indication of any fluorescence in my samples even in
the positive control. I don't know what wrong with it.  Does anybody have
this experience in examination of the GUS expression directly from the
leaves through the fluorescene IN MICROTITER PLATE.


     Any suggestion  will be helpful, Thank you


Regina