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 g

Na2SeO3 5 H2O 120.0 g

Na2WO4 2 H2O 30.0 g

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 121C 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 121C for 15 min

 

Updated 23.06.2021