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Medium Detail Information

Medium No.
Methanogenium Medium
 Trace element solution*10ml
 Vitamin solution**10ml
 Sodium acetate1g
 Bacto Yeast Extract (Difco)2g
 Trypticase Peptone (BBL)2g
 Distilled water1L
  Mix ingredients except NaHCO3, Na2S·9H2O and cysteine-HCl, and adjust pH to 6.5. Bring to a boil for 5-10 seconds, then cool under a stream of H2/CO2 (80/20) and add NaHCO3 to the medium. After autoclaving under a H2/CO2 atmosphere, stand the medium overnight. Separately autoclave cysteine-HCl and Na2S·9H2O as 5% solutions under a N2 atmosphere. Prior to inoculation, aseptically and anaerobically add the cysteine-HCl and Na2S·9H2O solutions. Pressurize the inoculated bottles to 200 kPa with H2/CO2.
  *Trace element solution
   Nitrilotriacetic acid1.5g
   Distilled water1L
  Dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH. Then add minerals. Adjust final pH to 7.0.
  **Vitamin solution
   Folic acid2mg
   Nicotinic acid5mg
   p-Aminobenzoic acid1mg
   Vitamin B120.01mg
   Distilled water1L