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Main sol. 579

Internal Solution-ID: S1244

Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
Fe(III) citrate 13.70 g 13.551 55.323
NH4Cl 1.50 g 1.484 27.738
NaH2PO4 0.60 g 0.593 4.947
KCl 0.10 g 0.099 1.327
Na-acetate 2.50 g 2.473 30.144
Modified Wolin's mineral solution 10.00 ml - -
Na2WO4 x 2 H2O
(0.1% w/v)
0.25 ml 2.5e-4 7.5e-4
NaHCO3 2.50 g 2.473 29.436
Wolin's vitamin solution (10x) 1.00 ml - -
Distilled water 1000.00 ml - -
1 First dissolve ferric citrate in water by heating and adjust to pH 6.0, then add other medium ingredients except bicarbonate and vitamins. Sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic, then add and dissolve bicarbonate and adjust pH to 6.8. Thereafter, dispense medium under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add vitamins to the sterile medium from an anoxic stock solution prepared under 100% N2 gas and sterilized by filtration. Adjust pH of complete medium to 6.7 - 7.0, if necessary.
2 Note: Use 5 - 10% (v/v) inoculum.
579 GEOBACTER MEDIUM