Medium data

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478   MODIFIED MMJHS MEDIUM FOR GO25

NaCl  20.0     g
K2HPO4  0.9     g
KH2PO4  0.7     g
CaCl2·2H2O  0.8     g
NH4Cl  0.25    g
NaNO3  1.0     g
MgSO4·7H2O  4.0     g
MgCl2·6H2O  3.0     g
KCl  0.33    g
NiCl2·6H2O  1.0     mg
Na2SeO3·5H2O  1.0     mg
Fe2(SO4)3·xH2O  0.01    g
H2WO4  1.0     mg
Na2S2O3·5H2O  2.0     g
Trace mineral solution (see Medium No. 368)  10.0     ml
Trace vitamins (see Medium No. 197)  1.0     ml
NaHCO3  2.0     g
Sulfur (powder)  10.0     g
Resazurin  1.0     mg
Distilled water  1.0     L
Mix ingredients except Trace vitamins, NaHCO3 and sulfur, adjust pH to 6.8 and autoclave. Filter-sterilize Trace vitamins and 5% NaHCO3 solution and add to the medium. Steam sulfur for 3 hr on each of 3 successive days. Distribute the medium into culture vessels containing appropriate amount of sulfur (e.g., 20 ml of medium in 120 ml serum bottles containing 0.2 g sulfur) under a stream of H2-CO2 (4:1, v/v) gas mixture and seal with butyl rubber stoppers. Pressurize the inoculated bottle to 200 kPa H2-CO2 (4:1, v/v).