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Main sol. 749

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Internal Solution-ID: S1535

Equipment needed: Filter Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2SO4 4.00 g 3.956 27.855
Na2HPO4 0.20 g 0.198 1.394
NH4Cl 0.25 g 0.247 4.623
MgCl2 x 6 H2O 0.40 g 0.396 1.946
KCl 0.50 g 0.495 6.634
CaCl2 x 2 H2O 0.15 g 0.148 1.009
Modified Wolin's mineral solution 10.00 ml - -
Yeast extract 0.50 g 0.495 -
Na-DL-lactate 2.50 g 2.473 22.067
Sodium resazurin
(0.1% w/v) 		0.50 ml 4.9e-4 0.002
NaHCO3 1.30 g 1.286 15.307
Wolin's vitamin solution (10x) 1.00 ml - -
Na-thioglycolate 0.20 g 0.198 1.734
L-Ascorbic acid 0.20 g 0.198 1.123
Distilled water 1000.00 ml - -
1 Dissolve ingredients except bicarbonate, vitamins, thioglycolate and ascorbic acid, then sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Dispense medium under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add vitamins, thioglycolate and ascorbic acid from anoxic stock solutions prepared under 100% N2 gas and sterilized by filtration and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Prior to use adjust pH of complete medium to 7.0, if necessary.
749 THERMODESULFOVIBRIO YELLOWSTONII MEDIUM