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Medium Detail Information

Medium No.
Medium for strain C/L2
 Trace element mixture*1ml
 Selenite-tungstate solution**1ml
 Vitamin mixture***1ml
 Vitamin B12 solution****1ml
 Bicarbonate solution (1.0 M)*****30ml
 Sulfide solution******24ml
 Distilled water1L
  Mix ingredients except vitamin mixture, vitamin B12 solution, bicarbonate solution and sulfide solution, and adjust pH to about 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. Aseptically and anaerobically add vitamin mixture, vitamin B12 solution, bicarbonate solution and sulfide solution using sterile syringes, and the N2/CO2 (80/20) mixed gas is enclosed with the vessels. Adjust pH to 7.2- 7.8 using compact pH meter. An aqueous solution of sodium lactate (final concentration, 20 mM) is added as the electron donor and carbon source before inoculation. Addition of yeast extract (0.5 - 1 g/L) greatly improves the growth of the strain.
  *The trace element mixture
   HCl (25%)12.5ml
   Distilled water987ml
  Sterilized by autoclaving.
  **The selenite-tungstate solution
   distilled water1L
  Sterilized by autoclaving.
  ***The vitamin mixture
   4-Aminobenzoic acid4mg
   Nicotinic acid10mg
   Calcium D(+)-panthothenate5mg
   Pyridoxine dihydrochloride15mg
   Distilled water100ml
  Sterilized by filtration.
  ****The vitamin B12 solution
   Distilled water100ml
  Sterilized by filtration.
  *****The bicarbonate solution (1.0 M)
   Distilled water1L
  Sterilized by filtration.
  ******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.