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Main sol. 684

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Internal Solution-ID: S1432

Equipment needed: Filter Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2HPO4 x 2 H2O 0.53 g 0.528 3.722
KH2PO4 0.41 g 0.409 3.004
NH4Cl 0.30 g 0.299 5.592
CaCl2 x 2 H2O 0.11 g 0.11 0.746
MgCl2 x 6 H2O 0.10 g 0.1 0.49
NaCl 0.30 g 0.299 5.118
Trace element solution SL-10 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
Yeast extract 0.20 g 0.199 -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
Na2CO3 1.50 g 1.496 14.11
Na2-fumarate 3.20 g 3.19 19.936
Wolin's vitamin solution (10x) 1.00 ml - -
Na2S x 9 H2O 0.50 g 0.499 2.076
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except carbonate, vitamins, fumarate and sulfide) 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. After autoclaving complete the medium by adding vitamins, fumarate and sulfide 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 vitamins and fumarate are sterilized by filtration. Adjust pH of the complete medium to 7.0 - 7.2. After inoculation pressurize culture vials to 0.7 bar overpressure with sterile 80% N2 and 20% CO2 gas mixture.
2 Note: A white precipitate forms in this medium after autoclaving, which has however no negative effect on growth.
684 SYNTROPHOBACTER (MPOB) MEDIUM