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Main sol. 509a

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Internal Solution-ID: S1123

Equipment needed: Gassing station Hungate tubes Autoclave Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 962 ml - -
Solution B 20 ml - -
Solution C 1 ml - -
Solution D 10 ml - -
Solution E 10 ml - -
1 Solution A is sparged with 80% N2 and 20% CO2 gas mixture to reach a pH below 6 (at least 30 -45 min), 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 atmosphere. Solutions C, E and F are autoclaved under 100% N2 gas atmosphere. Solution D is prepared under 100% N2 gas atmosphere and sterilized by filtration. To complete the medium appropriate amounts of solutions B to F are added to the sterile solution A in the sequence as indicated. Final pH of the medium should be 6.8 - 7.0.
509a SPIROCHAETA RI 19.B1 MEDIUM