Thermococcus siculi Grote et al.
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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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Grote R, et al. Thermococcus siculi sp. nov., a novel hyperthermophilic archaeon isolated from a deep-sea hydrothermal vent at the Mid-Okinawa Trough. Extremophiles 3: 55-62, 1999. PubMed: 10086845