Protocol for catalase and Oxidase test for bacterial cells
1. Catalase Test:
Procedure:
1. Transfer a small amount of bacterial colony to a surface of clean, dry glass slide using a loop or sterile wooden stick
2. Place a drop of 3% H2O2 on to the slide and mix.
3. A positive result is the rapid evolution of oxygen (within 5-10 sec.) as evidenced by bubbling.
4. A negative result is no bubbles or only a few scattered bubbles.
5. Dispose of your slide in the biohazard glass disposal container.
Reagent:
1. 3% H2O2
-Take 1 ml 30-35% H2O2 and add 9 ml D/w.
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| Catalse test |
2. Oxidase Test:
Procedure: (Wet Filter Method)
1. A strip of filter paper is soaked with a little freshly made 1% solution of tertramethyl-p-phenylene-diamine dihydrochloride reagent.
2. A speck of culture is rubbed on it with a platinum loop.
3. A positive reaction is indicated by an intense deep-purple hue, appearing within 5-10 seconds, a “delayed positive” reaction by colouration in 10-60 seconds, and a negative reaction by absence of colouration or by colouration later than 60 seconds.
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| Oxidase test |
Procedure: (Dry Filter Method)
1. Strip of Whatman’s No. 1 filter paper are soaked in a freshly prepared 1% solution of tertramethyl-p-phenylene-diamine dihydrochloride.
2. After draining for about 30 seconds, the strips are freeze dried and stored in a dark bottle tightly sealed with a screw cap.
3. For use, a strip is removed, laid in a petri dish and moistened with distilled water.
4. The colony to be tested is picked up with a platinum loop and smeared over the moist area.
5. A positive reaction is indicated by an intense deep-purple hue, appearing within 5-10 seconds, a “delayed positive” reaction by colouration in 10-60 seconds, and a negative reaction by absence of colouration or by colouration later than 60 seconds.
Reagents: 1% tetramethyl-p-phenylene-diamine dihydrochloride
-1 g tetramethyl-p-phenylene-diamine dihydrochloride and add 100 ml d/w
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