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Main sol. 1062

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Internal Solution-ID: S2152

Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.20 g 0.198 1.454
NH4Cl 0.25 g 0.247 4.623
NaCl 1.00 g 0.989 16.925
MgCl2 x 6 H2O 0.40 g 0.396 1.946
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 - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 4.9e-4 0.002
Na2CO3 1.50 g 1.484 13.999
FeSO4 x 7 H2O 22.00 mg 0.022 0.078
Seven vitamins solution 1.00 ml - -
Yeast extract 2.00 g 1.978 -
Na-pyruvate 4.50 g 4.451 40.448
Na2-fumarate 6.50 g 6.429 40.174
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except carbonate, ferrous iron sulfate, vitamins and organic substrates), 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. Add ferrous iron sulfate (dissolved in 0.1 N H2SO4), vitamins, pyruvate, fumarate and yeast extract 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 atmosphere. Stock solutions of ferrous iron sulfate, vitamins, pyruvate and fumarate should be sterilized by filtration. Adjust pH of the complete medium to 7.5, if necessary.
2 Note: Addition of 10 - 20 mg sodium dithionite per litre (e.g. from 5% (w/v) solution, freshly prepared under N2 and filter-sterilized) may stimulate growth at the beginning.
1062 DESULFITOBACTERIUM (PCE II) MEDIUM