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Medium Detail Information

Medium No.
Methanobacterium Medium
  KH2PO4 0.75 g
  K2HPO4 0.75 g
  NH4Cl 1 g
  MgCl2·6H2O 0.36 g
  Bacto Yeast Extract (Difco) 0.1 g
  Polypeptone 0.1 g
  Trace elements solution* 10 ml
  Vitamin solution** 10 ml
  Resazurin 1 mg
  NaHCO3 4 g
  Cysteine-HCl 0.5 g
  Na2S·9H2O 0.5 g
  Distilled water 1 L
    Mix ingredients except for vitamin solution, cysteine-HCl and Na2S·9H2O, and autoclave under H2/CO2 (80/20) condition. Separately autoclave cysteine-HCl and Na2S·9H2O as a 5% solution under a N2 atmosphere. Aseptically and anaerobically add filter-sterile vitamin solution, and cysteine-HCl and Na2S·9H2O solutions, and pressurize inoculated bottles to 150 kPa with H2/CO2.
    *Trace elements solution
      Nitrilotriacetic acid (NTA) 4.1 g
      FeCl3·6H2O 0.36 g
      MnCl2·4H2O 0.09 g
      CoCl2·6H2O 0.15 g
      CaCl2·2H2O 0.018 g
      ZnCl2 0.09 g
      H3BO3 0.017 g
      Na2MoO4·2H2O 0.009 g
      NiCl2·6H2O 0.018 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