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Main sol. 730

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Internal Solution-ID: S1479

Equipment needed: Gassing station Serum bottle Autoclave Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 950 ml - -
Solution B 30 ml - -
Solution C 1 ml - -
Solution D 10 ml - -
Solution E 10 ml - -
1 Sparge solution A with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic, then dispense under same gas atmosphere into anoxic serum vials to 10% volume (e.g., 10 ml medium in 100 ml bottle) and autoclave. Solution B is autoclaved separately after sparging with 80% N2 and 20% CO2 gas mixture. Solution C and D are prepared under 100% N2 gas atmosphere and sterilized by filtration. Solution E is autoclaved under 100% N2 gas atmosphere. To complete the medium add appropriate amounts of solutions B to E to the sterile solution A in the sequence as indicated.
2 Adjust pH of complete medium to 7.2, if necessary.
3 Note: Cultures are very sensitive to the nitrite produced during growth. Thus, incubate this strain ony a few hours at one time (8-12 h) and thereafter check growth by microscopy. Immediately stop the incubation at 85°C, when the medium begins to show some turbidity.
730 FERROGLOBUS PLACIDUS MEDIUM