Composition per liter:

NaCl ……19.45g

Sulfur ……10.0g

MgCl₂……8.8g

Peptone……5.0g

Na₂SO₃……3.24g

CaCl₂……1.8g

Yeast extract ……1.0g

KCl……0.55g

NaHCO₃ ……0.16g

Ferric citrate……0.1g

KBr……0.08g

SrCl₂……0.03g

H₃BO₃ ……0.02g

Na₂HPO₄ ……8.0mg

Na₂SiO₃……4.0mg

NaF……2.4mg

NH₄NO₃ ……1.6mg

pH 7.6 ± 0.2 at 25°C

Preparation of Sulfur: Autoclave sulfur for 15 min at 0 psi pressure–100°C on three successive days.

Preparation of Medium: Prepare anaerobically under a gas phase of 80% N₂ + 10% CO₂ + 10% H₂.

Add components, except sulfur, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat while stirring and bring to boiling.

Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50–55°C. Aseptically add 10.0g of sterile sulfur.

Mix thoroughly. Aseptically and anaerobically, under a gas phase of 80% N₂ + 10% CO₂ + 10% H₂, distribute into sterile tubes.

Use: For the cultivation and maintenance of Thermococcus litoralis.