Two biochemical tests were conducted to validate bacterial identification. First, was the test to validate oxidase positive bacteria, where an oxidase strip was touched to a loopful of bacteria. The other test was the microbact 12A strip, where a bacterial suspension was made by picking 1 to 3 isolated colonies from culture plate re-suspended in 2.5 milliliters of sterile saline. After opening the microbsct strip, 4 drops of bacterial suspension were added to each of the 12 wells. Three wells( 1,2,3) were overlaid with mineral oil using a sterile Pasteur pipette, resealed and incubated for 24 hours. After the 24 hours, 2 drops of Indole reagent was added to the eighth well and evaluated within 2 minutes. In well 10, 1 drop each of VPI and VPII reagent were added and evaluated after 15 minutes. In well 12, 1 drop of TDA reagent was added and results recorded immediately. The reaction in each well was recorded as either positive or negative by comparing to a color chart.
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