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Lab 7 Notebook
Title: AA 07 Secondary Assays for Bacterial Characterization Objective: To profile bacterial population based on additional biochemical approaches to better determine growth/metabolic characteristics Procedure:
- Indole Test: Test
o Using a sterile loop, pick an isolated colony from BAP o Smear Directly onto the reaction area of the slide o Examine test area for color change within 20 seconds
- Indole Test: Test
o Using sterile loop pick an isolated colony from BAP o Inoculate bacteria in media broth o Grow at 37C overnight
- TSI Test
o Use a sterile loop to inoculate slant from liquid culture o Press loop into agar slant o While removing loop, streak slanted side of agar with loop o Grow at 37C overnight o (next day) examine samples
- API Test
o Fill all tubes with bacteria culture
• Fill CIT, VP, and GEL entirely
• Add mineral oil to ADH, LDC, ODC, H25, and URE= anaerobic
o Add 5ml distilled water onto tray, add API strip and cover with lid o Incubate at 37C overnight o (next day) examine samples
o Pseudomonas: negative for indole o Salmonella: negative for indole
- TSI Test results
o Negative control: remained red (native color), has pH indicator o E. Coli: gram- bacteria, solid yellow on slant, able to ferment all 3 sugars, produces gas o Salmonella sample: gram – bacteria, yellow to black, produces hydrogen sulfide gas, ferments all 3 sugars
- API Test results
