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Medium No.
Methanosarcina acetivorans Medium
 Bacto Yeast Extract (Difco)1g
 Trace elements solution*10ml
 Vitamin solution**10ml
 Distilled water1L
  Mix ingredients except methanol, vitamin solution, cysteine-HCl, and Na2S·9H2O. Flush with N2/CO2, and adjust pH to 7.0 with HCl. Dispense the medium into suitable culture vessels under a stream of N2/CO2 (80/20) and seal with butyl rubber stoppers. Separately autoclave cysteine-HCl and Na2S·9H2O (5% solution) under a N2 atmosphere. Sterilize methanol and vitamin solution by filtration. Prior to inoculation, aseptically and anaerobically add vitamin, cysteine-HCl and Na2S·9H2O solutions.
  *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