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Medium Detail Information

Medium No.	1081
Medium	MMJHS medium
Composition
NaNO3 1 g   Na2S2O3·5H2O 1 g   NaHCO3 1 g   Sulfur (powder) 3 g   Balch's vitamin solution* 10 ml   MJ synthetic seawater** 990 ml   pH 6.5       Mix ingredients except for vitamin, sulfur and bicarbonate. Dispense the medium into suitable culture vessels under a stream of H2/CO2 (80/20), seal with butyl rubber stoppers, and autoclave at 121°C for 15 min. After cooling to room temperature, the pH is adjusted to 6.5 by adding of sterile pH-adjusting agent with a syringe without opening the vessel. Sulfur is separately sterilized by autoclaving for 60 min at 105°C on each of 3 successive days. Sterilize vitamin and bicarbonate solutions by filtration. Prior to inoculation, aseptically and anaerobically add the vitamin, sulfur and bicarbonate to the medium, and pressurize the vessels to 350 kPa with H2/CO2.                   *Balch's vitamin solution       Biotin 2 mg       Folic acid 2 mg       Pyridoxine-HCl 10 mg       Thiamine-HCl 5 mg       Riboflavin 5 mg       Nicotinic acid 5 mg       Ca-pantothenate 5 mg       Vitamin B12 0.1 mg       p-Aminobenzoic acid 5 mg       Lipoic acid 5 mg       Distilled water 1 L                   **MJ synthetic seawater       NaCl 30 g       K2HPO4 0.14 g       CaCl2·2H2O 0.14 g       NH4Cl 0.25 g       MgSO4·7H2O 3.4 g       MgCl2·6H2O 4.18 g       KCl 0.33 g       NiCl2·6H2O 0.5 mg       Na2SeO3·5H2O 0.5 mg       Fe(NH4)2(SO4)2·6H2O 0.01 g       Balch's trace mineral solution*** 10 ml       Distilled water 990 ml                   ***Balch's trace mineral solution       Nitrilotriacetic acid (NTA) 1.5 g       MgSO4·7H2O 3 g       MnSO4·2H2O 0.5 g       NaCl 1 g       FeSO4·7H2O 0.1 g       CoSO4·7H2O 0.18 g       CaCl2·2H2O 0.1 g       ZnSO4·7H2O 0.18 g       CuSO4·5H2O 0.01 g       KAl(SO4)2·12H2O 0.02 g       H3BO3 0.01 g       Na2MoO4·2H2O 0.01 g       NiCl2·6H2O 0.025 g       Na2SeO3·5H2O 0.3 mg       Distilled water 1 L     First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Final pH 7.0 (with KOH).
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