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Medium Detail Information

Medium No.
Marine H2-using Methanogen Medium 
 KP7 buffer*5ml
 Bacto Yeast Extract (Difco)0.02g
 Sodium acetate0.8g
 Trace elements solution**2ml
 Vitamin solution***2ml
 Distilled water1L
 pH unadjusted 
  Mix ingredients except KP7 buffer, vitamin solution, Na2CO3, Na2S·9H2O and cysteine-HCl. Dispense the medium into suitable culture vessels under a stream of H2/CO2 (80/20) and seal with butyl rubber stoppers. Separately autoclave KP7 buffer, Na2S·9H2O (5% solution) and cysteine-HCl (5% solution) under a N2 atmosphere. Sterilize vitamin and 10% Na2CO3 solutions by filtration. Prior to inoculation, aseptically and anaerobically add KP7 buffer, vitamin, Na2CO3 and Na2S·9H2O solutions. Pressurize the inoculated vessels to 150 kPa with H2/CO2.
  *KP7 buffer
   Distilled water1L
  Autoclaved under N2.
  **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

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