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Main sol. 212

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Internal Solution-ID: S445

Equipment needed: Gassing station Filter Hungate tubes Autoclave Centrifuge
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 942 ml - -
Solution B 30 ml - -
Solution C 10 ml - -
Solution D 1 ml - -
Solution E 10 ml - -
Solution F 10 ml - -
1 Add and dissolve ingredients of solution A and sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under the same gas atmosphere into anoxic Hungate-type tubes and autoclave. Solution B is prepared under 80% N2 and 20% CO2 gas atmosphere and autoclaved. Solutions C, E and F are autoclaved under 100% N2 gas atmosphere. Solution D is prepared under 100% N2 gas 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. Adjust pH of complete medium to 7.2, if necessary.
2 Note: Some cultures are shipped in semi-solid medium which stimulates growth at the beginning. For agar stabs 3.00 g/l agar are added to the complete medium from a sterile anoxic stock solution (2% w/v). Upon receipt add anoxically 1 - 2 ml of the recommended freshly prepared liquid medium to the agar tube and incubate for 3 - 5 days . After incubation transfer 0.5 ml of the resulting cell suspension in the liquid phase to tubes with liquid medium.
212 SYNTHROPHOMONAS MEDIUM