303. AMMONIFEX MEDIUM

KH₂PO₄ 0.34 g

NH₄Cl 0.34 g

CaCl₂ × 2 H₂O 0.05 g

KCl 0.34 g

Trace elements solution (see below) 1.0 ml

Vitamin solution (see below) 1.0 ml

NaHCO₃ 1,5 g

Na₂S × 7-9 H₂O 0.7 g

Na₂S₂O₃O × 5 H₂O 0.5 g

NaCl 25.0 g

MgCl₂ × 6 H₂O 0.33 g

Na-formate 2.0 g

pH 7.0

Trace element solution:

(NH₄)₂Fe(SO₄)₂ × 6 H₂O (Mohr's salt) 784.0 mg

CoCl₂ × H₂O 238.0 mg

(NH₄)₂Ni(SO₄)₂ × 6 H₂O 395.0 mg

Na₂MoO₄ × 2 H₂O 24.0 mg

Na₂WO₄ × 2 H₂O 33.0 mg

ZnSO₄ × 7 H₂O 144.0 mg

CuCl₂ × 2 H₂O 2.0 mg

Na₂SeO₄ 94.0 mg

H₃BO₃ 6.0 mg

MnCl₂ × 4 H₂O 99.0 mg

Vitamin solution:

Biotin 20.0 mg

Folic acid 20.0 mg

Pyridoxine 100.0 mg

Riboflavin 50.0 mg

Pantotenoic acid 50.0 mg

p-Aminobenzoic acid 50.0 mg

Thiamine-HCl 50.0 mg

Nicotinic acid 50.0 mg

Vitamin B₁₂ 1.0 mg

Lipoic (thioctic) acid 50.0 mg

Distilled water 1000.0 ml

Dissolve ingredients except Na-formate, Na₂S₂O₃, vitamins and sulfide. Adjust pH to 3.5 with H₂SO₄ and sparge medium with 100% CO₂ gas for 30 – 45 min to make it anoxic. Add Na-formate, Na₂S₂O₃, vitamins (sterilized by filtration) and sulfide from sterile anoxic stock solutions prepared under 100% N₂ gas