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Main sol. 507b

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Internal Solution-ID: S1114

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
Trace element solution SL-4 10.00 ml - -
Na2CO3 2.50 g 2.473 23.331
Na-pyruvate 1.00 g 0.989 8.988
Yeast extract 0.20 g 0.198 -
Wolin's vitamin solution (10x) 1.00 ml - -
Distilled water 1000.00 ml - -
1 Dissolve ingredients except carbonate, pyruvate, yeast extract, and vitamins, then sparge medium with 100% CO2 gas for 30 - 45 min to make it anoxic. Add carbonate and adjust pH to 6.0 - 6.5, then dispense medium under 100% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add pyruvate, yeast extract and vitamins from sterile anoxic stock solutions prepared under 100% N2 gas atmosphere. Stock solutions of pyruvate and vitamins are sterilized by filtration. After completing the medium adjust pH to 6.5 with a sterile anoxic stock solution of 1 N NaOH.
507b CARBOXYDOCELLA MANGANICA MEDIUM