232. Carboxydothermus ferrireducens medium

KH₂PO₄ 0.33 g

NH₄Cl 0.33 g

KCl 0.33 g

MgCl₂ × 6 H₂O 0.33 g

CaCl₂ × 2 H₂O 0.33 g

NaHCO₃ 2.0 g

Glycerol (87%) 3.0 ml

Vitamin solution (see below) 10.0 ml

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

NiCl₂ × 6 H₂O 200.0 µg

Na₂SeO₃ × 5 H₂O 120.0 µg

Na₂WO₄ × 2 H₂O 30.0 µg 

Yeast extract 1.0 g

Na₂-9,10-anthraquinone-2,6-disulfonate (Sigma A9706) 8.25 g

Distilled water 1000.0 ml

Vitamin solution:

Biotin 2.0 mg

Folic acid 2.0 mg

Pyridoxine 10.0 mg

Riboflavin 5.0 mg

Pantotenoic acid 5.0 mg

p-Aminobenzoic acid 5.0 mg

Thiamine-HCl 5.0 mg

Nicotinic acid 5.0 mg

Vitamin B₁₂ 0.1 mg

Lipoic acid 5.0 mg

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

Dissolve ingredients (except CaCl₂ × 2 H₂O, NaHCO₃, and vitamins), boil medium for some minutes to dissolve the anthraquinone, then cool under 80% N₂ and 20% CO₂ gas atmosphere to room temperature. Add solid NaHCO₃ and adjust medium pH to 6.8 with NaOH. Dispense medium in tubes or bottles under same gas. Autoclave at 121°C for 15 min. Before use, add CaCl₂ × 2 H₂O and vitamins from anoxic, sterile stock solutions. Vitamin solution is filter sterilized. CaCl₂ × 2 H₂O and trace element solution Autoclave at 121°C for 15 min

 

Updated 23.06.2021