Oxalobacter formigenes Allison et al.
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ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.
2. Aseptically transfer 0.5 ml of #1514 broth to the vial and rehydrate the pellet. Transfer the cell suspension back into the broth tube. Inoculate a plate of a non-selective medium such as Tryptic Soy, Nutrient, or blood agar with 0.1 ml of the cell suspension.
3. Seal the tube with a rubber stopper and incubate anaerobically at 20-25oC. Incubate the plate(s) aerobically as a purity check.
4. After two or three days, growth should be evident through out the broth. Once growth has been established the culture should be transferred to fresh broth every 24 to 48 hours.
5. This culture is very sensitive to oxygen therefore steps should be taken to avoid exposure to air. When the culture exhibits good growth it will remain viable for up to 1 week if stored at 4oC under anaerobic conditions.
· Tubes of media are placed under a gassing cannula system hooked to a source of oxygen-free gas.
· All transfers are performed while the test tubes are on the cannula system with a gentle stream of oxygen-free gas flowing through the system.
· As the test tubes are removed from the cannula system, each is sealed with butyl rubber stopper thus maintaining the anaerobic headspace.
· 100% carbon dioxide is typically employed as the oxygen-free gas source.
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