362. GEOALKALIBACTER MEDIUM

NH₄Cl 0.5 g

KCl 0.2 g

MgCl₂ × 6 H₂O 0.1 g

KH₂PO₄ 0.2 g

NaCl 1.0 g

Yeast extract 0.1 g

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

Selenite-tungstate solution (see below) 1.0 ml

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

Na₂CO₃ 3.0 g

NaHCO₃ 10.0 g

Sulfur, powdered 10.0 g

Na-acetate × 3 H₂O 2.5 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

Selenite-tungstate solution:

NaOH 0.5 g

Na₂SeO₃ × 5 H₂O 3.0 mg

Na₂WO₄ × 2 H₂O 4.0 mg

Distilled water 1000.0 ml

Dissolve ingredients (except carbonates, sulfur and acetate), then sparge medium with 100% N₂ gas for 30 – 45 min to make it anoxic. Add solid carbonate and bicarbonate, adjust pH to 9.0 - 9.2, dispense under 100% N₂ gas atmosphere into anoxic Hungate type tubes or serum vials containing already the appropriate amount of sulfur. Sterilize medium by heating cultivation vessels in a water bath to 90 – 100°C for 1 – 2 hours on each of 3 successive days. Add acetate from a sterile anoxic stock solution prepared under 100% N₂ gas.

 

Updated 1.02.2024