272. ECTOTHIORHODOSPIRA MAGNA MEDIUM

KH₂PO₄ 0.5 g

NaCl 30.0 g

NH₄Cl 0.5 g

MgCl₂ × 6 H₂O 0.2 g

Na-acetate 1.0 g

Na₂S₂O₃ 0.5 g

Na₂S × 9 H₂O 0.5 g

NaHCO₃ 5.0 g

Na₂CO₃ 5.0 g

Yeast extract 0.1 g

Vitamin B₁₂ 10.0 mkg

Trace elements solution (see below) 1.0 ml

Distilled water 1000.0 ml

pH 9.0-9.5

Prepare the medium without the NaHCO₃, Na₂S × 9 H₂O, Na₂CO₃ and vitamin B₁₂, under a nitrogen atmosphere.

Autoclave at 121°C for 15 min and add the vitamin B12 from a filter-sterilized stock solution and the NaHCO₃ (autoclaved in sealed, half full vessels), Na₂S × 9 H₂O, Na₂CO₃ from autoclaved at 121°C for 15 min stock solutions. The final pH of the medium should be 9.0-9.5.

Trace elements solution:

EDTA 5.0 g

FeSO₄ × 7 H₂O 2.2 g

ZnSO₄ × 7 H₂O 0.1 g

MnCl₂ × 4 H₂O 0.03 g

H₃BO₃ 0.03 g

CoCl₂ × 6 H₂O 0.2 g

CuCl₂ × 2 H₂O 0.03 g

NiCl₂ × 6 H₂O 0.03 g

Na₂MoO₄ × 2 H₂O 0.03 g

Distilled water 1000.0 ml

pH 3.0-4.0