182. DESULFOBACTERIUM MEDIUM

Solution 1:

Na₂SO₄ 3.0 g

KH₂PO₄ 0.2 g

NH₄Cl 0.3 g

NaCl 21.0 g

MgCl₂ × 6 H₂O 3.0 g

KCl 0.5 g

CaCl₂ × 2 H₂O 0.15 g

Resazurin 1.0 mg

Distilled water 930.0 ml

Solution 2:

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

Solution 3:

Vitamin solution (see below) 10.0 ml

Solution 4:

NaHCO₃ 2.5 g

Distilled water 50.0 ml

Solution 5:

Na₂SeO₃ × 5 H₂O (3 mg in 1000.0 ml 0.01 M NaOH) 1.0 ml

Solution 6:

Na₂S × 9 H₂O 0.4 g

Distilled water 10.0 ml

Solition 7:

Substrate - depending on the species of bacteria:

25% Na-acetate 10.0 ml or indole 0.3 g

NaCl 2.1 g

MgCl₂ × 6 H₂O 0.3 g

Distilled water 100.0 ml

Phenol 40.0 mg

or Na-benzoate 400.0 mg

Distilled water 4.0 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:

p-Aminobenzoic acid 4.0 mg

D(+)-Biotin 1.0 mg

Thiamine-HCl 10.0 mg

Distilled water 100.0 ml

pH 7.1-7.4

Solution 1 is prepared and autoclaved anaerobically under 80% N₂ + 20% CO₂ at 121°C for 15 min. Solutions 2, 4, 5 and 6 are gassed with N₂ and autoclaved separately at 121°C for 15 min. Solution 3 (gassed with N₂ + CO₂) and solution 7 (gassed with N₂) are filter-sterilized.

 

Updated 27.05.2023