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Medium Detail Information

Medium No.
955
Medium
Hydrogen-using methanogen medium
Composition
 KCl0.355g
 MgCl2·6H2O4g
 MgSO4·7H2O3.45g
 NH4Cl0.25g
 CaCl2·2H2O0.14g
 KH2PO40.14g
 Fe(NH4)2(SO4)2·6H2O2mg
 Sodium acetate0.82g
 Bacto Yeast Extract (Difco)2g
 Hipolypepton (Nihon seiyaku)2g
 Trace elements solution*10ml
 Vitamin solution**10ml
 NaHCO35g
 Resazurin1mg
 Cysteine-HCl0.5g
 Na2S·9H2O0.5g
 Distilled water1L
  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)12.8g
   FeCl3·6H2O1.35g
   MnCl2·4H2O0.1g
   CoCl2·6H2O0.024g
   CaCl2·2H2O0.1g
   ZnCl20.1g
   CuCl2·2H2O0.025g
   H3BO30.01g
   Na2MoO4·2H2O0.024g
   NaCl1g
   NiCl2·6H2O0.12g
   Na2SeO40.004g
   Na2WO40.004g
   Distilled water1L
  First dissolve nitrilotriacetic acid and adjust pH to 6.5 with NaOH, and then add minerals. Final pH 7.0.
       
  **Vitamin solution
   Biotin2mg
   Folic acid2mg
   Pyridoxine-HCl10mg
   Thiamine-HCl5mg
   Riboflavin5mg
   Nicotinic acid5mg
   Ca-pantothenate5mg
   p-Aminobenzoic acid1mg
   Vitamin B120.01mg
   Distilled water1L
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