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Medium Detail Information

Medium No.
 Sodium acetate1g
 Sodium succinate1g
 Yeast extract0.1g
 Vitamin mixture*1ml
 Trace element solution**1ml
 Neutralized Na2S·9H2O0.5g
 Distilled water1L
 Agar (if needed)15g
 pH 7.3-7.5 
  Mix ingredients (except vitamin mixture) and adjust pH to 7.3-7.5. Dispense the medium into suitable culture vessels under a stream of nitrogen gas, seal with butyl rubber stoppers, and autoclave at 121°C for 15 min. Separately autoclave neutralized sulfide solution*** under a N2 atmosphere. Sterilize vitamin mixture by filtration. Sterile syringes are used to inoculate and remove samples. Prior to inoculation, aseptically and anaerobically add vitamin mixture and neutralized sulfide solution.
  *Vitamin mixture
   p-Aminobenzoic acid30mg
   Vitamin B125mg
   Distilled water100ml
  **Trace element solution
   Distilled water1L
  ***Neutralized sulfide solution
   Distilled water100ml
  The sulfide solution is prepared in a vessel sealed with a butyl rubber stopper. Autoclave at 121℃ for 15 min. under a N2 atmosphere. After cooling to room temperature the pH is adjusted to about 7.3 by adding of sterile 2N H2SO4 drop-wise with a syringe without opening the bottle. Appearance of yellow colour indicates the pH is about 8. The vessel should be shaken continuously to avoid precipitation of elemental sulfur. The final solution should be transparent yellow.