Stenoxybacter acetivorans Wertz and Breznak

1. Sterilize the top of the all test tubes by spraying them with 70% ethanol and then flaming the tops.
2. If needed, exchange the gas in each test tube for 80% N₂ 20% CO₂ (do not over pressurize; keep the pressure below 5 PSI).  Add 1 to 2 ml of air to each tube using a 1 or 5 ml syringe.
3. Thaw the frozen vial at room temperature.
4. For inoculation, use a 1.0 ml syringe tipped with 22-gauge needle, and withdraw the cell suspension from the vial and transfer it to the broth tube.  Transfer 0.5 ml of the culture to an additional tube of #2678 broth.   Plate 0.1 ml of the inoculated culture onto a non-selective medium and incubate aerobically at 30°C. Use 0.1 ml of the inoculated culture to inoculate a nonselective aerobic broth.  Incubate the broth tubes at 30°C.
5. Growth should be detected in the #2678 broth within 4 to 7 days.  There should be no growth detected on the aerobic plate or in the aerobic broth.

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