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Medium Detail Information

Medium No.
1013
Medium
Modified Pfennig's Medium II (green sulfur bacteria)
Composition
 CaCl2·2H2O0.25g
 KH2PO40.34g
 NH4Cl0.34g
 KCl0.34g
 MgSO4·7H2O1g
 Vitamin B12 (0.002%) solution1ml
 Trace element solution SL-10B*1ml
 NaHCO31.5g
 Na2S·9H2O0.6g
 Resazurin0.5mg
 Distilled water998ml
 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%)7.7ml
   FeSO4·7H2O1.5g
   ZnCl270mg
   MnCl2·4H2O100mg
   H3BO3300mg
   CoCl2·6H2O190mg
   CuCl2·2H2O2mg
   NiCl2·6H2O24mg
   Na2MoO4·2H2O36mg
   Distilled water1L
       
  **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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