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Medium No.	1138
Medium	Green Sulfur Bacterium Medium (1% NaCl)
Composition
KH2PO4 0.5 g   MgSO4·7H2O 0.5 g   CaCl2·2H2O 0.05 g   KCl 0.025 g   NH4Cl 0.5 g   Ammonium acetate 0.5 g   NaCl 10 g   Na2S2O3 (if needed) 0.5 g   Ferric citrate solution (0.1%)* 5 ml   Vitamin B12 solution (0.001%) 2 ml   Trace element solution SL-6** 2 ml   Acetate solution*** 10 ml   NaHCO3 1.5 g   Na2S·9H2O (neutralized)**** 0.6 g   Distilled water 1 L     Mix ingredients except vitamin B12, acetate solution, NaHCO3 and Na2S·9H2O, distribute the medium into culture vessels, seal with butyl rubber stoppers, and autoclave (121°C, 15 min) under a N2/CO2 atmosphere (80/20). Aseptically add vitamin B12, acetate, NaHCO3 (30 g/L) and neutralized Na2S·9H2O solutions.                   *Ferric citrate     Soluble not water.                   **Trace element solution SL-6       ZnSO4·7H2O 0.1 g       MnCl2·4H2O 0.03 g       H3BO3 0.3 g       CoCl2·6H2O 0.2 g       CuCl2·2H2O 0.01 g       NiCl2·6H2O 0.02 g       Na2MoO4·2H2O 0.03 g       Distilled water 1 L                   ***Acetate solution       Ammonium acetate 25 g       Magnesium acetate 25 g       Fructose 25 g       Distilled water 1 L     Mix ingredients, adjust pH to 7.2, and autoclave under a N2 atmosphere.                   ****Na2S·9H2O (neutralized)       Na2S·9H2O 15 g       Distilled water 1 L     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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