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Main sol. 1451

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Internal Solution-ID: S2983

Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.33 g 0.329 2.418
NH4Cl 0.33 g 0.329 6.151
KCl 0.33 g 0.329 4.413
NaCl 18.00 g 17.946 307.087
CaCl2 x 2 H2O 0.33 g 0.329 2.238
MgCl2 x 6 H2O 4.00 g 3.988 19.616
Yeast extract 0.20 g 0.199 -
Trace element solution SL-10 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
Ferric citrate monohydrate 2.50 g 2.493 9.479
Na2CO3 1.00 g 0.997 9.407
Wolin's vitamin solution (10x) 1.00 ml - -
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except ferric citrate, carbonate and vitamins) and 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 ferric citrate (dissolve by boiling and adjust pH to 6.5 - 7.0) and vitamins 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 mixture. Sterilize vitamins by filtration. Adjust pH of the complete medium to 6.5, if necessary.
1451 DEFERRISOMA MEDIUM