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Main sol. 480

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Internal Solution-ID: S5981

Equipment needed: Hungate tubes Filter Gassing station Autoclave Water bath
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 0.33 g 0.329 6.157
CaCl2 x 2 H2O 0.33 g 0.329 2.24
MgCl2 x 6 H2O 0.33 g 0.329 1.62
KCl 0.33 g 0.329 4.418
KH2PO4 0.33 g 0.329 2.42
Trace element solution SL-10 1.00 ml - -
Yeast extract 0.10 g 0.1 -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
Sulfur
(powder) 		10.00 g 9.98 311.195
Na-acetate 5.00 g 4.99 60.829
NaHCO3 2.50 g 2.495 29.7
Wolin's vitamin solution (10x) 1.00 ml - -
DL-Dithiothreitol 0.30 g 0.299 1.94
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except sulfur, acetate, bicarbonate, vitamins, and dithiothreitol) and sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Adjust pH to 5.9, dispense under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials containing already the appropriate amount of sulfur and sterilize by heating for 2-3 hours in a boiling water bath on each of 3 successive days. Add acetate, vitamins, and dithiothreitol from sterile anoxic stock solutions prepared under 100% N2 gas and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Solutions of vitamins and dithiothreitol are sterilized by filtration. Prior to using adjust the pH of the complete medium to 6.8 - 7.0, if necessary.
480 DESULFURELLA MEDIUM