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Medium Detail Information

Medium No.
1005
Medium
Artificial seawater medium for strain JS_SRB250Lac
Composition
  KH2PO4 0.2 g
  NH4Cl 0.25 g
  NaCl 26 g
  MgCl2·6H2O 11.2 g
  KCl 0.7 g
  CaCl2·2H2O 1.4 g
  Na2SO4 4 g
  KBr 0.1 g
  NaHCO3 2.5 g
  Sodium lactate 2.3 g
  Resazurin 0.5 g
  Trace element solution SL-10* 1 ml
  Selenite-tungstate solution** 1 ml
  Vitamin solution*** 1 ml
  Sulfide solution**** 10 ml
  Distilled water 1 L
 
    Dissolve ingredients except dihydrogenphosphate, ammonium chloride, bicarbonate, lactate, vitamins and sulfide, and autoclave it in a cultivation device for anaerobes. After sterilization and cooling under a nitrogen atmosphere, add dihydrogenphosphate, ammonium chloride, bicarbonate, lactate, vitamins and sulfide from sterile stock solutions. Adjust the final pH of the medium to 7.2-7.4. Dispense the medium under N2/CO2 (80/20) gas mixture in culture vessels.
             
    *Trace element solution SL-10
      HCl (25%; 7.7 M) 10 ml
      FeCl2·4H2O 1.5 g
      ZnCl2 7 mg
      MnCl2·4H2O 100 mg
      H3BO3 6 mg
      CoCl2·6H2O 190 mg
      CuCl2·2H2O 2 mg
      NiCl2·6H2O 24 mg
      Na2MoO4·2H2O 36 mg
      Distilled water 990 ml
             
    **Selenite-tungstate solution
      NaOH 0.5 g
      Na2SeO3·5H2O 3 mg
      Na2WO4·2H2O 4 mg
      distilled water 1 L
 
    Sterilized by autoclaving.
             
    ***Vitamin solution
      Biotin 2 mg
      Folic acid 2 mg
      Pyridoxine-HCl 10 mg
      Thiamine-HCl 5 mg
      Riboflavin 5 mg
      Nicotinic acid 5 mg
      Ca-pantothenate 5 mg
      Vitamin B12 0.1 mg
      p-Aminobenzoic acid 5 mg
      Lipoic acid 5 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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