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Main sol. 1098

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Internal Solution-ID: S2214

Equipment needed: Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
Sea Salt
(SIGMA) 		35.00 g 34.93 -
NH4Cl 0.10 g 0.1 1.866
KH2PO4 0.05 g 0.05 0.367
Na-acetate 1.60 g 1.597 19.465
Yeast extract 0.10 g 0.1 -
Wolfe's mineral elixir 1.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
NaHCO3 2.00 g 1.996 23.76
2-Mercaptoethanesulfonic acid
(coenzyme M) 		0.14 g 0.14 0.983
Seven vitamins solution 1.00 ml - -
Na2S2O3 x 5 H2O 2.50 g 2.495 10.053
DL-Dithiothreitol 0.30 g 0.299 1.94
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except bicarbonate, coenzyme M, vitamins, thiosulfate, and dithiothreitol), then sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under the same gas atmosphere into anoxic serum vials (e.g., Balch-type tubes) to 30% of their volume and autoclave. Add coenzyme M, vitamins, thiosulfate, and dithiothreitol from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas atmosphere. The vitamin and dithiothreitol solutions should be sterilized by filtration. Adjust the pH of the complete medium to 6.5.
2 After inoculation pressurize culture vessels to 1.5 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture.
1098 ARCHAEOGLOBUS INFECTUS MEDIUM