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Main sol. 395

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Internal Solution-ID: S806

Equipment needed: Hungate tubes Filter Gassing station Water bath
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 0.33 g 0.329 6.157
KH2PO4 0.33 g 0.329 2.42
KCl 0.33 g 0.329 4.418
CaCl2 x 2 H2O 0.44 g 0.439 2.987
MgCl2 x 6 H2O 0.70 g 0.699 3.436
NaCl 0.50 g 0.499 8.539
Trace element solution SL-10 1.00 ml - -
Yeast extract
(OXOID) 		0.20 g 0.2 -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
Sulfur
(powdered) 		10.00 g 9.98 311.195
D-Glucose 2.50 g 2.495 13.849
Wolin's vitamin solution (10x) 1.00 ml - -
Na2S x 9 H2O 0.50 g 0.499 2.078
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except sulfur, glucose, vitamins and sulfide), then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Adjust pH to 6.2 - 6.4 and dispense medium under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials containing already the appropriate amount of sulfur. Sterilize medium by heating cultivation vessels in a boiling water bath for 2 - 3 hours on each of 3 successive days. After sterilization add glucose, vitamins and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas atmosphere. Vitamins are sterilized by filtration. Adjust pH of complete medium to 6.5, if necessary.
395 DESULFUROCOCCUS MEDIUM