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Medium Detail Information

Medium No.
870
Medium
Medium for strain C/L2
Composition
 Na2SO44g
 KH2PO40.2g
 NH4Cl0.25g
 NaCl20g
 MgCl2·6H2O3g
 KCl0.5g
 CaCl2·2H2O0.15g
 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
   FeSO4·7H2O2100mg
   H3BO330mg
   MnCl2·4H2O100mg
   CoCl2·6H2O190mg
   NiCl2·6H2O24mg
   CuCl2·2H2O2mg
   ZnSO4·7H2O144mg
   Na2MoO4·2H2O36mg
   Distilled water987ml
  Sterilized by autoclaving.
       
  **The selenite-tungstate solution
   NaOH0.4g
   Na2SeO·5H2O6mg
   Na2WO4·2H2O8mg
   distilled water1L
  Sterilized by autoclaving.
       
  ***The vitamin mixture
   4-Aminobenzoic acid4mg
   D(+)-Biotin1mg
   Nicotinic acid10mg
   Calcium D(+)-panthothenate5mg
   Pyridoxine dihydrochloride15mg
   Thiamine-HCl10mg
   Distilled water100ml
  Sterilized by filtration.
       
  ****The vitamin B12 solution
   Cyanocobalamin5mg
   Distilled water100ml
  Sterilized by filtration.
       
  *****The bicarbonate solution (1.0 M)
   NaHCO384g
   Distilled water1L
  Sterilized by filtration.
       
  ******Neutralized sulfide solution
   Na2S·9H2O1.5g
   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.
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