Bacterial V.P. assay
Summary
This experiment is from the official website of the Fourth Military Medical University
Operation method
Bacterial V.P. assay
Materials and Instruments
Escherichia coli Enterobacter aerogenes Proteus vulgaris Bacillus subtilis
NaOH solution Creatine Methyl red reagent Indole reagent Ether Bromomethylphenol violet indicator
Ultra-clean bench Constant temperature incubator Autoclave Test tubes Pipettes Duchenne cannulae Glucose peptone water medium Peptone water medium Sugar fermentation medium
Move
1. Marking of test tubes
Take a test tube containing glucose peptone culture solution and mark the cultured bacteria as required.
2. Inoculation culture
Aseptically inoculate a small amount of bacterial moss into the above corresponding test tubes, the blank control tube is not inoculated, placed in a constant temperature of 37 ℃, culture 24 ~ 48h.
3. Observation record
Take out the above test tubes, oscillate for 2min, take 5 empty test tubes and mark the bacteria name accordingly, add 3~5ml of culture solution in the above corresponding tubes, then add 10~20 drops of 40% NaOH solution and add about 0.5~1mg of microcreatine, oscillate the tubes so as to dissolve the oxygen in the air, and then keep warm at 37℃ constant temperature for 15~30min, if the culture solution shows red color, then it will be recorded as V If the culture solution shows red color, it will be recorded as positive reaction of V.P. test (indicated by "+"); if it does not show red color, it will be recorded as negative reaction of V.P. test (indicated by "-").
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