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Main sol. 507a

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Internal Solution-ID: S1113

Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
KCl 0.33 g 0.326 4.378
MgCl2 x 6 H2O 0.52 g 0.514 2.53
CaCl2 x 2 H2O 0.29 g 0.287 1.951
NH4Cl 0.33 g 0.326 6.102
KH2PO4 0.33 g 0.326 2.399
Na2-9,10-anthraquinone-2,6-disulfonate
(TCI A0308) 		8.25 g 8.16 -
Trace element solution SL-4 10.00 ml - -
NaHCO3 1.00 g 0.989 11.774
Yeast extract 0.20 g 0.198 -
Wolin's vitamin solution (10x) 1.00 ml - -
Distilled water 1000.00 ml - -
1 Dissolve ingredients except bicarbonate, yeast extract, vitamins and sulfide, then sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% of their volume and autoclave. Add yeast extract, vitamins (sterilized by filtration) and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas atmosphere and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. After completing the medium adjust pH to 6.8 - 7.0.
2 Inoculated vessels are pressurized with sterile carbon monoxide gas to 1 bar overpressure.
507a CARBOXYDOTHERMUS SIDEROPHILUS MEDIUM