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Medium Detail Information

Medium No.
 Fumarate solution (1.0 M)*10ml
 Trace element mixture**1ml
 Selenite-tungstate solution***1ml
 Bicarbonate solution (1.0 M)****30ml
 Vitamin solution*****1ml
 Thiamine solution******1ml
 Vitamin B12 solution*******1ml
 Sulfide solution (1.0 M)********1.5ml
 Distilled water953.5ml
 pH 7.0-7.3 
  Mix ingredients except for fumarate, bicarbonate, vitamin, thiamine, cyanocobalamin and sulfide solutions. Dispense the medium into suitable culture vessels under a stream of the N2/CO2 (80/20) mixed gas, seal with butyl rubber stoppers, and autoclave at 121°C for 15 min. Aseptically and anaerobically add other ingredients using sterile syringes. Adjust pH to 7.0-7.3 using compact pH meter. Moreover, prior to inoculation, aseptically and anaerobically add the fumarate solution (final concentration, 10 mM).
  *Fumarate solution (1.0 M)
   Disodium fumarate1.6g
   Distilled water10ml
  Sterilized by filtration and stored under N2 gas.
  **Trace element mixture
   HCl (25%; 7.7 M)12.5ml
   Distilled water987ml
  Sterilized by autoclaving.
  ***Selenite-tungstate solution
   Distilled water1L
  Sterilized by autoclaving.
  ****Bicarbonate solution (1.0 M)
   Distilled water100ml
  Saturated with CO2 gas. Tightly sealed in bottles filled with CO2 gas and then sterilized by autoclaving.
  *****Vitamine solution
   p-Aminobenzoic acid4mg
   Nicotinic acid10mg
   Sodium phosphate buffer (10 mM; pH7.1)100ml
  Sterilized by filtration.
  ******Thiamine solution
   Sodium phosphate buffer (25 mM; pH 3.4)100ml
  Sterilized by filtration.
  *******Vitamin B12 solution
   Distilled water100ml
  Sterilized by filtration.
  ********Sulfide solution (1.0 M)
   Distilled water100ml
  Sterilized by filtration and stored under N2 gas.