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Medium Detail Information

Medium No.	869
Medium	Medium for strain C/G2
Composition
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   Dithionite (ca.) 20 mg   Distilled water 1 L     Mix ingredients except vitamin mixture, vitamin B12 solution, bicarbonate solution and dithionite, 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 dithionite 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 glutamate (final concentration, 10 mM) and sodium thiosulfate (final concentration, 20 mM) are added before inoculation. Addition of yeast extract (0.5 - 1 g/L) greatly improves the growth of the strain.                   *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.                   **Selenite-tungstate solution       NaOH 0.4 g       Na2SeO3·5H2O 6 mg       Na2WO4·2H2O 8 mg       Distilled water 1 L       Sterilized by autoclaving.                   ***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.                   ****Vitamin B12 solution       Cyanocobalamin 5 mg       Distilled water 100 ml       Sterilized by filtration.                   *****Bicarbonate solution (1.0 M)       NaHCO3 84 g       Distilled water 1 L       Sterilized by filtration.
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