Read Microsoft Word - GUS staining (histochemical) text version

Histochemical staining for GUS activity

Judelson protocol Staining solution 50 mM NaPO4, pH 7.0 0.1% Triton X-100 0.1% X-Gluc (bromochloroindoyl-b-glucuronide) in dimethyl formamide store this in the dark! When it turns red, it has gone bad. optional reagents to increase sensitivity (oxidation catalysts) 5 mM potassium ferricyanide (K3Fe(CN)6) 5 mM potassium ferrocyanide (K4Fe(CN)6) Procedure 1. Staining fungal mycelia Place a tuft of mycelia in a microfuge tube or microtiter plate. Add staining solution to cover Place at 37C Color will appear in 5-60 minutes; can stain overnight if desired 2. Staining plant material Place entire leaf (or cut off small pieces from leaf) in staining solution Vacuum infiltrate (suck vacuum and release; may need to repeat until air removed from leaf). Incubate at 37C overnight Decolorize with 95% ethanol; rinse several times; do final rinse in water before viewing under microscope

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Microsoft Word - GUS staining (histochemical)

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