Smittium simulatum Lichtw. et Arenas
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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.
1. To thaw a frozen ampoule, place in a 2530 °C water bath, until just thawed (approximately 5 minutes). Immerse the ampoule just sufficient to cover the frozen material. Do not agitate the ampoule.
2. Immediately after thawing, wipe down ampoule with 70% ethanol and aseptically transfer 10 microliter (or any amount desired up to all) of the content onto a plate or broth with medium recommended.
3. Incubate the inoculum/strain at the temperature and conditions recommended.
4. Inspect for growth of the inoculum/strain regularly. The sign of viability is noticeable typically after 1-2 days of incubation. However, the time necessary for significant growth will vary from strain to strain.
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Lichtwardt RW, Mrenas JM. Trichomycetes in aquatic insects from southern Chile. Mycologia 88: 844-857, 1996.