Thermococcus siculi Grote 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. Using a single tube of #2104 broth (5 to 6 ml) under anaerobic conditions, withdraw approximately 0.5 to
1.0 ml with a Pasteur or 1.0 ml pipette. Re-hydrate the entire pellet.
3. Aseptically transfer this aliquot back into the broth tube. Mix well. Take 0.25 ml of this tube and transfer it to a second tube. Mix well.
4. After 24 Hours of incubation, turbidity should be observed.
5. Transfers can now be made to other tubes of broth, slants, and/or plates.
Anaerobic conditions for transfer may be obtained by either of the following:
· Use of an anaerobic gas chamber, or
· Placement of test tubes under a gassing cannula system hooked to anaerobic gas or.
· Exchanging the headspace in stoppered culture tubes for the appropriate anaerobic gas
Anaerobic conditions for incubation may be obtained by any of the following:
· Loose screw‑caps on test tubes in anaerobic chamber,
· Loose screw‑caps on test tubes in an activated anaerobic Gas Pak jar, or
· Use of sterile butyl rubber stoppers on test tubes so that an anaerobic gas headspace is retained.
growth is enhanced when tubes are pressurized (20-40 psi).
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