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Main sol. 1243

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Internal Solution-ID: S2466

Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
Ferric citrate monohydrate 13.70 g 13.551 51.532
NaCl 1.00 g 0.989 16.925
MgCl2 x 6 H2O 0.40 g 0.396 1.946
KH2PO4 0.20 g 0.198 1.454
NH4Cl 0.25 g 0.247 4.623
KCl 0.50 g 0.495 6.634
CaCl2 x 2 H2O 0.15 g 0.148 1.009
Modified Wolin's mineral solution 10.00 ml - -
NaHCO3 2.50 g 2.473 29.436
Na-pyruvate 2.50 g 2.473 22.471
Wolin's vitamin solution (10x) 1.00 ml - -
Na2S x 9 H2O 0.10 g 0.099 0.412
Distilled water 1000.00 ml - -
1 First dissolve ferric citrate by heating the water. After cooling to room temperature adjust the pH to 6.0 and dissolve the other ingredients (except bicarbonate, vitamins, pyruvate and sulfide). Sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Before inoculation add vitamins, pyruvate and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Stock solutions of pyruvate and vitamins should be sterilized by filtration. Adjust pH of complete medium to 6.8 - 7.2, if necessary.
1243 PARADESULFITOBACTERIUM MEDIUM