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Main sol. 1074

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Internal Solution-ID: S2168

Equipment needed: Autoclave Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
Sea Salt
(SIGMA) 		25.00 g 24.925 -
Na2SO4 3.50 g 3.49 24.567
NH4Cl 0.25 g 0.249 4.66
KH2PO4 0.20 g 0.199 1.465
Wolfe's mineral elixir 1.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
Na2CO3 1.50 g 1.496 14.11
Yeast extract 0.05 g 0.05 -
Na-pyruvate 1.00 g 0.997 9.06
Wolin's vitamin solution (10x) 1.00 ml - -
n-Decane 1.00 ml - -
Na2S x 9 H2O 0.10 g 0.1 0.415
L-Cysteine HCl x H2O 0.10 g 0.1 0.568
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except carbonate, yeast extract, pyruvate, vitamins, n-decane and reducing agents) and sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic serum vials (e.g., Balch-type tubes or serum bottles) and autoclave. Add yeast extract, pyruvate, vitamins, n-decane and reducing agents from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Solutions of vitamins and pyruvate should be sterilized by filtration. Adjust pH of complete medium to 7.2 - 7.4.
2 Note: Use 20% (v/v) as inoculum and incubate vials in an inverted position.
1074 DESULFOGLAEBA MEDIUM