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Main sol. 386a

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Internal Solution-ID: S6347

Equipment needed: Gassing station Hungate tubes Autoclave Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 967 ml - -
Solution B 30 ml - -
Solution C 1 ml - -
Solution D 10 ml - -
Sulfur
(powdered) 		10 g 9.921 309.343
1 Solution A is sparged with 80% N2 and 20% CO2 gas mixture for 30 – 45 min to make it anoxic, then distributed under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclaved. Solution B is autoclaved separately under 80% N2 and 20% CO2 gas mixture. Solutions C and D are prepared under a 100% N2 gas atmosphere and sterilized by filtration. To complete the medium solutions B to D are added to the sterile solution A in the sequence as indicated. Before inoculation, appropriate amounts of sterile sulfur are added to the vials with the medium. Sulfur is sterilized by steaming for 3 hours on each of 3 successive days (see medium 35) and added aseptically to the sterile medium while retaining anoxic conditions. The final pH of the medium should be at 7.0.