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Main sol. 1040

Internal Solution-ID: S2108

Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
Sea Salt
(SIGMA)
35.0 g 34.93 -
Wolfe's mineral elixir 1.0 ml - -
Yeast extract 1.0 g 0.998 -
Sodium resazurin
(0.1% w/v)
0.5 ml 5.0e-4 0.002
Na2CO3 1.0 g 0.998 9.416
Na-DL-lactate 2.5 g 2.495 22.265
Wolin's vitamin solution (10x) 1.0 ml - -
Na2S x 9 H2O 0.3 g 0.299 1.247
Distilled water 1000.0 ml - -
1 Dissolve ingredients (except carbonate, lactate, 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. After autoclaving add lactate, vitamins and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile stock solution prepared under 80% N2 and 20% CO2 gas mixture. The vitamin solution should be sterilized by filtration. Adjust pH of the complete medium to 7.0-7.2, if necessary.
1040 MARIDESULFOVIBRIO MEDIUM