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Main sol. 770

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Internal Solution-ID: S1568

Equipment needed: Gassing station Serum bottle Autoclave Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 970 ml - -
Solution B 10 ml - -
Solution C 10 ml - -
Solution D 5 ml - -
Solution E 10 ml - -
1 Dissolve ingredients of solution A except carbonate, sparge with 100% N2 gas for 30 - 45 min to make it anoxic, then add carbonate and adjust pH to 9.0 - 9.3. Dispense under 100% N2 gas atmosphere into anoxic Hungate-type tubes or serum vials (e.g., 30 ml medium per 100 ml bottle) and autoclave. Solutions B and C are autoclaved separately under 100% N2 gas atmosphere. Solutions D and E are prepared under 100% N2 gas and sterilized by filtration. To complete the medium add appropriate amounts of solutions B to E to the sterile solution A in the sequence as indicated. Adjust pH of complete medium to 7.5.
2 Note: Addition of 10 - 20 mg sodium dithionite per liter (e.g. from 5% (w/v) solution, freshly prepared under N2 and filter-sterilized) may stimulate growth at the beginning.
3 For transfers use 10% (v/v) inoculum.
770 SULFOPHOBOCOCCUS ZILLIGII MEDIUM