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).