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Main sol. 796

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Internal Solution-ID: S1624

Equipment needed: Autoclave Filter Gassing station Serum bottle
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 18.00 g 17.946 307.087
KCl 0.33 g 0.329 4.413
MgCl2 x 6 H2O 7.15 g 7.129 35.064
NH4Cl 0.25 g 0.249 4.66
CaCl2 x 2 H2O 0.14 g 0.14 0.949
K2HPO4 x 3 H2O 0.18 g 0.179 1.03
Trace element solution 1.00 ml - -
Fe(NH4)2(SO4)2 x 6 H2O
(0.1% w/v) 		2.00 ml 0.002 -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
NaHCO3 4.00 g 3.988 47.473
Na2SO3 1.00 g 0.997 7.91
Na-acetate 1.00 g 0.997 12.154
Na2S x 9 H2O 0.50 g 0.499 2.076
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except bicarbonate, sulfite, acetate and sulfide), then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Thereafter, add and dissolve bicarbonate and equilibrate the medium with the 80% N2 and 20% CO2 gas mixture to achieve a medium pH of 7.0. Dispense medium under same gas atmosphere into serum vials (e.g., 25 ml per 100 ml bottle) and autoclave. After sterilization add sulfite, acetate and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. The stock solution of sulfite should be freshly prepared and sterilized by filtration. Prior to inoculation check pH of complete medium and adjust to 7.0, if necessary.
2 After inoculation, pressurize bottles to 1 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture.
796 ARCHAEOGLOBUS VENEFICUS MEDIUM