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Medium Detail Information

Medium No.
Methanosaeta thermophila medium
  NH4Cl 0.5 g
  K2HPO4 0.4 g
  MgCl2·6H2O 0.1 g
  CaCl2·2H2O 0.1 g
  Sodium acetate 3.3 g
  Coenzyme M 0.14 g
  NaHCO3 1 g
  Vitamin solution* 10 ml
  Trace element solution** 10 ml
  Resazurin 0.5 mg
  Na2S·9H2O 0.25 g
  Distilled water 1 L
    Mix ingredients except for CaCl2·2H2O, sodium acetate, coenzyme M, NaHCO3, vitamin solution and Na2S·9H2O, and autoclave under a N2/CO2 atmosphere (80/20). Concentrated CaCl2·2H2O, sodium acetate, coenzyme M and Na2S·9H2O solutions are sterilized separately under a N2 atmosphere in tightly closed vessels. Concentrated NaHCO3 solution and vitamin are filter-sterilized. Prior to inoculation, add CaCl2·2H2O, sodium acetate, coenzyme M, NaHCO3, vitamin and Na2S·9H2O solutions. Final pH is 6.5.
    *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
    **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
      KAl(SO4)2·12H2O 0.02 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.