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Medium Detail Information

Medium No.
 Yeast extract0.3g
 Disodium succinate1g
 Ammonium acetate0.5g
 Ferric citrate solution* (0.1%)5ml
 Vitamin B12 solution (0.01%)0.4ml
 Trace element solution SL-6**1ml
 Neutralized sulfide solution***5ml
 Distilled water1L
  Mix ingredients except ethanol, vitamin B12 solution and neutralized sulfide solution, and adjust pH to 6.8 with 6 N H2SO4. 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 ethanol and vitamin B12 by filtration. Sterile syringes are used to inoculate and remove samples. Prior to inoculation, aseptically and anaerobically add ethanol, vitamin B12 solution and neutralized sulfide solution.
  *Soluble in hot water.
  **Trace element solution SL-6
   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 2 N 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.