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Medium Detail Information

Medium No.
Hydrogen-using methanogen medium
 Sodium acetate0.82g
 Bacto Yeast Extract (Difco)2g
 Hipolypepton (Nihon seiyaku)2g
 Trace elements solution*10ml
 Vitamin solution**10ml
 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
   Distilled water1L
  First dissolve nitrilotriacetic acid and adjust pH to 6.5 with NaOH, and then add minerals. Final pH 7.0.
  **Vitamin solution
   Folic acid2mg
   Nicotinic acid5mg
   p-Aminobenzoic acid1mg
   Vitamin B120.01mg
   Distilled water1L