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Main sol. 508

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Internal Solution-ID: S1115

Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
KCl 0.33 g 0.326 4.374
MgCl2 x 6 H2O 0.52 g 0.514 2.527
CaCl2 x 2 H2O 0.29 g 0.287 1.949
NH4Cl 0.33 g 0.326 6.096
KH2PO4 0.33 g 0.326 2.396
Trace element solution SL-4 10.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 4.9e-4 0.002
Na2CO3 1.00 g 0.988 9.323
Na-pyruvate 2.50 g 2.47 22.449
Yeast extract 0.05 g 0.049 -
Seven vitamins solution 1.00 ml - -
Wolin's vitamin solution (10x) 1.00 ml - -
Na2S x 9 H2O 0.30 g 0.296 1.234
Distilled water 1000.00 ml - -
1 Dissolve ingredients except carbonate, pyruvate, yeast extract, vitamins and sulfide, then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 -45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add pyruvate, yeast extract, vitamins and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas atmosphere and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Stock solutions of pyruvate and vitamins are sterilized by filtration. After completing the medium adjust pH to 6.8-7.0 with a sterile anoxic stock solution of Na2CO3 (5% w/v), if necessary.
508 CARBOXYDOTHERMUS MEDIUM (HETEROTROPH)