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Medium Detail Information

Medium No.	955
Medium	Hydrogen-using methanogen medium
Composition
KCl 0.355 g   MgCl2·6H2O 4 g   MgSO4·7H2O 3.45 g   NH4Cl 0.25 g   CaCl2·2H2O 0.14 g   KH2PO4 0.14 g   Fe(NH4)2(SO4)2·6H2O 2 mg   Sodium acetate 0.82 g   Bacto Yeast Extract (Difco) 2 g   Hipolypepton (Nihon seiyaku) 2 g   Trace elements solution* 10 ml   Vitamin solution** 10 ml   NaHCO3 5 g   Resazurin 1 mg   Cysteine-HCl 0.5 g   Na2S·9H2O 0.5 g   Distilled water 1 L     Mix ingredients except vitamin solution, NaHCO3, cysteine-HCl and Na2S·9H2O, and adjust pH to 6.5 with NaOH. Dispense the medium into suitable culture vessels under a stream of H2/CO2 (80/20) and seal with butyl rubber stoppers. Separately autoclave cysteine-HCl and Na2S·9H2O as 5% solutions under a N2 atmosphere. Sterilize vitamin and 8% NaHCO3 solutions by filtration. Prior to inoculation, aseptically and anaerobically add vitamin, NaHCO3, cysteine-HCl and Na2S·9H2O solutions. Pressurize the inoculated vessels to 150 kPa with H2/CO2. Adjust pH to 6.8-7.0.                   *Trace elements solution       Nitrilotriacetic acid (NTA) 12.8 g       FeCl3·6H2O 1.35 g       MnCl2·4H2O 0.1 g       CoCl2·6H2O 0.024 g       CaCl2·2H2O 0.1 g       ZnCl2 0.1 g       CuCl2·2H2O 0.025 g       H3BO3 0.01 g       Na2MoO4·2H2O 0.024 g       NaCl 1 g       NiCl2·6H2O 0.12 g       Na2SeO4 0.004 g       Na2WO4 0.004 g       Distilled water 1 L     First dissolve nitrilotriacetic acid and adjust pH to 6.5 with NaOH, and then add minerals. Final pH 7.0.                   **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       p-Aminobenzoic acid 1 mg       Vitamin B12 0.01 mg       Distilled water 1 L
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