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Medium Detail Information

Medium No.	1013
Medium	Modified Pfennig's Medium II (green sulfur bacteria)
Composition
CaCl2·2H2O 0.25 g   KH2PO4 0.34 g   NH4Cl 0.34 g   KCl 0.34 g   MgSO4·7H2O 1 g   Vitamin B12 (0.002%) solution 1 ml   Trace element solution SL-10B* 1 ml   NaHCO3 1.5 g   Na2S·9H2O 0.6 g   Resazurin 0.5 mg   Distilled water 998 ml   pH 6.8       Dissolve ingredients except Vitamin B12 and sulfide, dispense in culture vessels, and autoclave under a N2/CO2 atmosphere (80/20). Add vitamin and sulfide** from sterile anaerobic stock solutions. The vitamin solution should be sterilized by filtration. Adjust the pH of the medium with sterile HCl or Na2CO3 solutions (2 mol/L each) to pH 6.8.                   *Trace element solution SL-10B       HCl (25%) 7.7 ml       FeSO4·7H2O 1.5 g       ZnCl2 70 mg       MnCl2·4H2O 100 mg       H3BO3 300 mg       CoCl2·6H2O 190 mg       CuCl2·2H2O 2 mg       NiCl2·6H2O 24 mg       Na2MoO4·2H2O 36 mg       Distilled water 1 L                   **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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