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Main sol. 334a

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Internal Solution-ID: S622

Equipment needed: Centrifuge Gassing station Serum bottle Autoclave Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 942 ml - -
Solution B 25 ml - -
Solution C 10 ml - -
Solution D 10 ml - -
Solution E 1 ml - -
Solution F 1 ml - -
Solution G 1 ml - -
Solution H 10 ml - -
1 Sparge solution A with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic, dispense under same gas atmosphere into anoxic serum vials or balch tubes (e.g., 20 ml medium in 50 ml bottles) and autoclave. Prior to inoculation complete the medium by adding sterile solutions C - H prepared under 100% N2 gas and solution B prepared under 80% N2 and 20% CO2 gas atmosphere. Solutions E, F and G are sterilized by filtration. Adjust the pH of the complete medium to 7.2.
2 Notes: Medium is only stable for a few weeks and has to be prepared freshly. Use 20% (v/v) inoculum. Sodium sulfide should be as pure as possible (use only clean crystals for the preparation of stock solutions). It has been noted that impurities of sulfide stock solutions can inhibit growth.
334a METHANOTHRIX SOEHNGENII MEDIUM