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Medium Detail Information

Medium No.	870
Medium	Medium for strain C/L2
Composition
Na2SO4 4 g   KH2PO4 0.2 g   NH4Cl 0.25 g   NaCl 20 g   MgCl2·6H2O 3 g   KCl 0.5 g   CaCl2·2H2O 0.15 g   Trace element mixture* 1 ml   Selenite-tungstate solution** 1 ml   Vitamin mixture*** 1 ml   Vitamin B12 solution**** 1 ml   Bicarbonate solution (1.0 M)***** 30 ml   Sulfide solution****** 24 ml   Distilled water 1 L     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%) 12.5 ml       FeSO4·7H2O 2100 mg       H3BO3 30 mg       MnCl2·4H2O 100 mg       CoCl2·6H2O 190 mg       NiCl2·6H2O 24 mg       CuCl2·2H2O 2 mg       ZnSO4·7H2O 144 mg       Na2MoO4·2H2O 36 mg       Distilled water 987 ml     Sterilized by autoclaving.                   **The selenite-tungstate solution       NaOH 0.4 g       Na2SeO·5H2O 6 mg       Na2WO4·2H2O 8 mg       distilled water 1 L     Sterilized by autoclaving.                   ***The vitamin mixture       4-Aminobenzoic acid 4 mg       D(+)-Biotin 1 mg       Nicotinic acid 10 mg       Calcium D(+)-panthothenate 5 mg       Pyridoxine dihydrochloride 15 mg       Thiamine-HCl 10 mg       Distilled water 100 ml     Sterilized by filtration.                   ****The vitamin B12 solution       Cyanocobalamin 5 mg       Distilled water 100 ml     Sterilized by filtration.                   *****The bicarbonate solution (1.0 M)       NaHCO3 84 g       Distilled water 1 L     Sterilized by filtration.                   ******Neutralized sulfide solution       Na2S·9H2O 1.5 g       Distilled water 100 ml     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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