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Medium Detail Information

Medium No.
1392
Medium
Modified Methanoculleus Medium
Composition
  NH4Cl 0.5 g
  MgSO4·7H2O 4.0 g
  CaCl2·2H2O 0.14 g
  K2HPO4 0.14 g
  KCl 0.1 g
  NaCl 6 g
  Fe(NH4)2(SO4)2·6H2O 2 mg
  (NH4)2Ni(SO4)2·6H2O 2 mg
  Vitamin solution* 10 ml
  Trace elements solution** 10 ml
  Se/W/AI solution*** 2 ml
  Glucose 5 g
  Resazurin 1 mg
  NaHCO3 2.5 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 N2/CO2 (80/20) condition. Separately autoclave cysteine-HCl and Na2S·9H2O as a 5% solution under a N2 atmosphere. Aseptically and anaerobically add cysteine-HCl and Na2S·9H2O solutions, and filter-sterile vitamin solution.
             
    *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
             
    **Trace element solution
      Nitrilotriacetic acid (NTA) 12.8 g
      FeCl3·6H2O 1.35 g
      MnCl2·4H2O 0.1 g
      CoCl2·6H2O 24 mg
      CaCl2·2H2O 0.1 g
      ZnCl2 0.1 g
      CuCl2·2H2O 25 mg
      H3BO3 10 mg
      Na2MoO4·2H2O 24 mg
      NaCl 1 g
      NiCl2·6H2O 0.12 g
      Distilled water 1 L
 
    First dissolve nitrilotriacetic acid and adjust pH to 6.5 with NaOH, then add minerals. Final pH 7.0.
             
    ***Se/W/AI solution
      Na2SeO3 4 mg
      Na2WO4 4 mg
      KAl(SO4)2·12H2O 20 mg
      Distilled water 1 L
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