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Problems with GUS batches

Melissa Melan melan at duq3.cc.duq.edu
Thu Aug 29 14:26:07 EST 1996


Hmmmm...*is* there a problem with X-Gluc from Rose?

I had problems with tissue staining a few months ago and assumed that 
it was due to my (and my students') lack of expertise and not the 
substrate.

I had an advanced undergraduate lab produce transgenic tobacco with 
pBI121.  We assayed tissue with p-nitrophenyl-Gluc and also did tissue 
staining with X-Gluc.  We got enzyme activity values for 20:20 plants 
with p-nitrophenyl-Gluc but tissue staining for only 3:20 plants 
(roots only; no staining in _any_ leaves).  The tissue staining was 
done using both vacuum infiltration and Triton-X 100 in the stain 
mixture.  The tissue staining was not restricted to only those plants 
with high enzyme activity.  I was (and still am) at a loss to explain 
this.  If anyone has any viable explanations for getting consistent 
tissue staining please pass them along.  Thanks!

======================================================================
Melissa Melan
Department of Biological Sciences
Duquesne University
Pittsburgh, PA  15282
(412) 396-5565



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