308. THERMOSIPHO AFFECTUS MEDIUM

Sea Salts (SIGMA) 30.0 g

Yeast extract (OXOID) 0.1 g

Trace element solution SL-10 (see below) 1.0 ml

Trypticase peptone 2.0 g

Na-resazurin solution (0.1% w/v) 0.5 ml

D-Glucose 2.0 g

Vitamin solution (see below) 10.0 ml

Na₂S × 9 H₂O 0.25 g

Distilled water 1000.0 ml

Trace element solution SL-10:

HCl (25%; 7.7 M) 10.0 ml

FeCl₂ × 4 H₂O 1.5 g

ZnCl₂ 70.0 mg

MnCl₂ × 4 H₂O 100.0 mg

H₃BO₃ 6.0 mg

CoCl₂ × 6 H₂O 190.0 mg

CuCl₂ × 2 H₂O 2.0 mg

NiCl₂ × 6 H₂O 24.0 mg

Na₂MoO₄ × 2 H₂O 36.0 mg

Distilled water 990.0 ml

Vitamin solution:

Biotin 2.0 mg

Folic acid 2.0 mg

Pyridoxine-HCl 10.0 mg

Thiamine-HCl × 2 H₂O 5.0 mg

Riboflavin 5.0 mg

Nicotinic acid 5.0 mg

D-Ca-pantothenate 5.0 mg

Vitamin B₁₂ 0.1 mg

p-Aminobenzoic acid 5.0 mg

Lipoic acid 5.0 mg

Distilled water 1000.0 ml

Dissolve ingredients except glucose, vitamins and sulfide, then sparge medium with 100% N₂ gas for 30-45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After sterilization add glucose, vitamins and sulfide from sterile anoxic stock solution prepared under 100% N₂ gas. The vitamin solution should be sterilized by filtration. Adjust pH of complete medium to 6.5–7.0, if necessary.

Updated 30.06.2021