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Main sol. 334

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Internal Solution-ID: S613

Equipment needed: Gassing station Serum bottle Autoclave Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 951 ml - -
Solution B 25 ml - -
Solution C 10 ml - -
Solution D 10 ml - -
Solution E 1 ml - -
Solution F 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 (e.g., 20 ml medium in 50 ml bottles) and autoclave. Prior to inoculation complete the medium by adding appropriate amounts of sterile solutions C, D, E and F prepared under 100% N2 gas and solution B prepared under 80% N2 and 20% CO2 gas atmosphere. Vitamins are sterilized by filtration. Adjust the pH of the complete medium to 7.2.
2 Note: Use 20% (v/v) inoculum.
334 METHANOTHRIX MEDIUM