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Histochemical GUS assay

时间:2006-07-24 14:10来源:Scienceboard.net 作者:admin
Thursday, December 04, 2003

Description
The histochemical assay to screen for expression of b-glucurodinase (gus) activity in genetically modified plants is carried out according to the method of Jefferson et al. (1987)

Procedure
For analysis, leaf tissue is incubated in reaction buffer containing 50 mM NaH2PO4 (pH 7), 0.01% Tween 20, 10 mM Na2EDTA and 0.3% (w/v) 5-bromo-4-chloro-3-indolyl glucuronide as a substrate for the enzyme. Tissue is incubated at 37篊 overnight and screened for blue staining indicating gus activity under a stereo-microscope. A short treatment with methanol is carried out before visual examination to intensify the blue staining.

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