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Main sol. 358a

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Internal Solution-ID: S698

Equipment needed: Serum bottle Gassing station Water bath Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
(NH4)2SO4 1.30 g 1.287 9.741
KH2PO4 0.28 g 0.277 2.037
MgSO4 x 7 H2O 0.25 g 0.248 1.004
CaCl2 x 2 H2O 0.07 g 0.069 0.471
FeCl3 x 6 H2O 0.02 g 0.02 0.073
Allen's trace element solution 10.00 ml - -
Sulfur
(powder) 		5.00 g 4.951 154.365
Yeast extract
(OXOID) 		0.50 g 0.495 -
Distilled water 1000.00 ml - -
1 Dissolve ingredients, except sulfur and yeast extract, bring medium to the boil, then cool to room temperature under 80% H2 and 20% CO2 gas mixture and adjust pH to 2.5 using 10 N H2SO4. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials (e.g., 20 ml medium in 100 ml serum bottles) containing already the appropriate amount of sulfur. For sterilization sealed bottles with medium are heated in a boiling water bath for 2 - 3 h on each of 3 successive days. Add yeast extract from a sterile anoxic stock solution prepared under 100% N2 gas atmosphere.
2 Pressurize inoculated bottles to 1 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture.
3 Note: Inoculate with 5% (w/v) culture. Incubate without shaking.
358a ACIDIANUS MEDIUM (ANAEROBIC)