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Medium Detail Information

Medium No.
Methanospirillum Medium
 Bacto Yeast Extract (Difco)2g
 Trypticase peptone (BBL)2g
 Sodium acetate2.5g
 Sodium formate2.5g
 Trace minerals*10ml
 Trace vitamins**10ml
 Distilled water1L
  Mix ingredients except NaHCO3, cysteine-HCl and Na2S·9H2O, and bring to a boil for 5-10 sec. Cool down under a stream of H2/CO2 (80/20), and add NaHCO3 to the medium. Separately autoclave cysteine-HCl and Na2S·9H2O as 5% solutions, respectively, under a N2 atmosphere. Prior to inoculation, aseptically and anaerobically add the cysteine-HCl and Na2S·9H2O solutions. Pressurize the inoculated vessels to 200 kPa with H2/CO2 (80/20).
  *Trace minerals
   Nitrilotriacetic acid1.5g
   Distilled water1L
  Dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH solution. Then proceed to add minerals. Adjust final pH to 7.0.
  ** Trace vitamins
   Folic acid2mg
   Nicotinic acid5mg
   Vitamin B120.1mg
   p-Aminobenzoic acid5mg
   Lipoic acid5mg
   Distilled water1L