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Main sol. 169b

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Internal Solution-ID: S295

Equipment needed: Autoclave Hungate tubes Gassing station Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 952 ml - -
Solution B 20 ml - -
Solution C 20 ml - -
Solution D 1 ml - -
Solution E 10 ml - -
1 Dissolve ingredients of solution A (except cysteine) and sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Add cysteine and adjust pH 7.2 with KOH, then distribute under the same gas atmosphere in anoxic Hungate-type tubes or serum vials and autoclave. Solution B is autoclaved separately under 80% N2 and 20% CO2 gas atmosphere, solution C is autoclaved under 100% N2 gas atmosphere and solutions D and E are sterilized by filtration under 100% N2 gas atmosphere. To complete the medium appropriate amounts of solutions B, C, D and E are added to the sterile solution A in the sequence as indicated. Final pH of the complete medium should be 7.5.
169b LEADBETTERA MEDIUM