Medium data

Search for medium no. = [811].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

811   THERMOCOCCALES RICH MEDIUM

PIPES  3.3     g
NaCl  23.0     g
MgCl2·6H2O  5.0     g
KCl  0.7     g
(NH4)2SO4  0.5     g
K2HPO4  0.05    g
KH2PO4  0.05    g
CaCl2·2H2O  0.02    g
NaBr  0.05    g
SrCl2·6H2O  0.01    g
Na2WO4·2H2O  3.3     mg
FeCl3·6H2O  6.8     mg
Yeast extract  1.0     g
Tryptone (BD-Difco)  4.0     g
Sulfur (powder)  10.0     g
Na2S·9H2O  0.5     g
Resazurin  1.0     mg
Distilled water  1.0     L
Mix ingredients except Na2S·9H2O, adjust pH to 6.8 and distribute the medium into culture vessels (e.g., 70 ml in 120 ml serum bottles). Bubble the medium with N2 gas for 30 min and seal with butyl rubber stoppers. Steam the medium for 3 hr on each of 3 successive days. Reduce the medium with sterile Na2S·9H2O solution (neutralized and autoclaved under a N2 atmosphere) and readjust pH to 6.8, if necessary.
Comment: For cultivation of JCM 16557, inoculated cultures should be pressurized to 20 to 100 MPa. Therefore, aspirate the culture into a plastic syringe with a 18G needle and stick the needle into a silicone rubber stopper. Put the assemblage of the syringe, needle and rubber stopper in a pressure vessel, pour water into the vessel and keep pressure inside between 20 and 100 MPa (at 90C).