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Dissolve compounds of solution A except sulfur and sparge it with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Distribute under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials that contain the appropriate amount of sulfur and autoclave at 110°C for 20 min. Solution B is autoclaved under 80% N2 and 20% CO2 gas atmosphere. Solution C is prepared under 100% N2 gas and sterilized by filtration. To complete the medium appropriate amounts of solutions B and C are added to the sterile solution A in the sequence as indicated. Adjust pH of complete medium to 7.5 - 8.0, if necessary.
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