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Main sol. 1028

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Internal Solution-ID: S2087

Equipment needed: Autoclave Hungate tubes Filter Gassing station Serum bottle
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 1.00 g 0.997 18.639
MgCl2 x 6 H2O 0.33 g 0.329 1.618
CaCl2 x 2 H2O 0.10 g 0.1 0.678
KCl 0.33 g 0.329 4.413
KH2PO4 0.50 g 0.499 3.663
Wolfe's mineral elixir 1.00 ml - -
Na-acetate 0.20 g 0.199 2.431
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
NaHCO3 0.50 g 0.499 5.934
Na2CO3 0.50 g 0.499 4.703
Na2S x 9 H2O 1.00 g 0.997 4.151
Yeast extract 0.20 g 0.199 -
Wolin's vitamin solution (10x) 2.00 ml - -
Distilled water 1000.00 ml - -
1 Dissolve ingredients except carbonates, sulfide, yeast extract and vitamins, then sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Add carbonates and sulfide to the medium, dispense under 100% carbon monoxide gas atmosphere into anoxic Hungate-type tubes or serum vials (e.g., 10 ml medium in 50 ml bottles) and autoclave. Prior to inoculation add yeast extract and vitamins (sterilized by filtration) from sterile anoxic stock solutions prepared under 100% N2 gas. If the medium is not completely reduced after autoclaving (remains pink) add 0.30 g/l sodium sulfide from a sterile anoxic stock solution prepared under 100% N2 gas atmosphere. Adjust pH of complete medium to 8.0 with a sterile anoxic solution of 1 N HCl.
1028 THERMINCOLA MEDIUM (CO)