quantitative lacZ measurements

Frederik [iso-8859-1] Börnke ricky_boernke at gmx.net
Tue Sep 12 03:42:14 EST 2000


Dear yeasters,

I am trying to quantifiy my two-hybrid positives using the ONPG
assay. I know that I have some strong ones, however, I obtain
obsolutely no signal. I even left the samples incubating over
night, but nothing. Since I have some clones which do not interact
I grew the cultures in trp/leu medium in order to have growth in
any case. I am aware of suboptimal reporter gene expression under
these conditions, however, at least with the strong interactors
I should see something, shouldn't I? Filterlifts work fine with
colonies grown on trp/leu plates. Is there anything else to 
consider? Are there any alternatives to quantify interaction?
Any hints appreciated.

Cheers
Ricky
 
*******************************************************************

Frederik Boernke
Research Group of  Molecular Plant Physiology
Institute for Plant Genetics and Crop Plant Research (IPK)
Corrensstr. 3
06466 Gatersleben
Tel.  039482 -5 321
Fax. 039482 -5 515
http://www.ipk-gatersleben.de





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