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Medium Detail Information

Medium No.
1321
Medium
Medium for Strain Pf12B
Composition
 NaCl20g
 MgCl2·6H2O3g
 CaCl2·2H2O0.15g
 Na2SO44g
 NH4Cl0.25g
 KH2PO40.2g
 KCl0.5g
 Sodium formate0.68g
 Yeast extract0.5g
 Resazurin1mg
 Trace element mixture*1ml
 Selenite-tungstate solution**1ml
 Bicarbonate solution (1.0 M)***30ml
 Vitamin solution****1ml
 Thiamine solution*****1ml
 Vitamin B12 solution******1ml
 Sulfide solution (1.0 M)*******1.5ml
 Distilled water963.5ml
 pH 7.0-7.3 
  Mix ingredients except for yeast extract, bicarbonate, vitamin, thiamine, cyanocobalamin and sulfide solutions. Dispense the medium into suitable culture vessels under a stream of the N2/CO2 (80/20) mixed gas, seal with butyl rubber stoppers, and autoclave at 121°C for 15 min. Aseptically and anaerobically add other ingredients using sterile syringes. Adjust pH to 7.0-7.3 using compact pH meter.
       
  *Trace element mixture
   HCl (25%; 7.7 M)12.5ml
   FeSO4·7H2O2100mg
   H3BO330mg
   MnCl2·4H2O100mg
   CoCl2·6H2O190mg
   NiCl2·6H2O24mg
   CuCl2·2H2O2mg
   ZnSO4·7H2O144mg
   Na2MoO4·2H2O36mg
   Distilled water987ml
  Sterilized by autoclaving.
       
  **Selenite-tungstate solution
   NaOH0.4g
   Na2SeO3·5H2O6mg
   Na2WO4·2H2O8mg
   Distilled water1L
  Sterilized by autoclaving.
       
  ***Bicarbonate solution (1.0 M)
   NaHCO38.4g
   Distilled water100ml
  Saturated with CO2 gas. Tightly sealed in bottles filled with CO2 gas and then sterilized by autoclaving.
       
  ****Vitamine solution
   p-Aminobenzoic acid4mg
   Biotin1mg
   Nicotinic acid10mg
   Ca-pantothenate5mg
   Pyridoxine-HCl15mg
   Sodium phosphate buffer (10 mM; pH7.1)100ml
  Sterilized by filtration.
       
  *****Thiamine solution
   Thiamine-HCl10mg
   Sodium phosphate buffer (25 mM; pH 3.4)100ml
  Sterilized by filtration.
       
  ******Vitamin B12 solution
   Cyanocobalamin5mg
   Distilled water100ml
  Sterilized by filtration.
       
  *******Sulfide solution (1.0 M)
   Na2S·9H2O24g
   Distilled water100ml
  Sterilized by filtration and stored under N2 gas.
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