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Medium Detail Information

Medium No.
Methanosaeta concilii medium
 Sodium acetate4.1g
 Trace element solution*10ml
 Vitamin solution**10ml
 Rumen fluid***10ml
 Distilled water1L
  Mix ingredients except for vitamin solution, rumen fluid, FeSO4, Na2CO3, cysteine-HCl and Na2S·9H2O, and autoclave under N2/CO2 (80/20) condition. A white precipitate that appears in the medium after autoclaving should dissolve again in about 2 days. Separately autoclave cysteine-HCl and Na2S·9H2O as a 5% solution under a N2 atmosphere. Vitamin and Na2CO3 (as a 10% solution) are sterilized by filtration. Aseptically and anaerobically add vitamin, rumen fluid, FeSO4, Na2CO3, cysteine-HCl and Na2S·9H2O. Adjust the pH of the medium to 7.3-7.5. Use 20% culture as an inoculum.
  *Trace element solution
   Nitrilotriacetic acid (NTA)4.1g
   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
  ***Rumen fluid:
  Remove solid particles from a fresh rumen fluid by filtration and centrifugation, and then autoclave under a N2 atmosphere. Stock at 4°C.
  FeSO4·7H2O is dissolved in 0.1 N H2SO4 as a 0.2% solution and then autoclaved under a N2 atmosphere.