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Main sol. 962a

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Internal Solution-ID: S1986

Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 0.33 g 0.326 6.102
KH2PO4 0.33 g 0.326 2.399
MgCl2 x 6 H2O 0.33 g 0.326 1.606
CaCl2 x 2 H2O 0.33 g 0.326 2.22
KCl 0.33 g 0.326 4.378
Yeast extract 0.05 g 0.049 -
Modified Wolin's mineral solution 10.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 4.9e-4 0.002
NaHCO3 0.70 g 0.692 8.242
Na2-fumarate 3.20 g 3.165 19.778
Meat extract
(OXOID) 		3.00 g 2.967 -
Wolin's vitamin solution (10x) 1.00 ml - -
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except bicarbonate, fumarate, meat extract and vitamins), then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add solid bicarbonate and adjust pH to 6.8 - 7.0. Dispense medium under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add fumarate, meat extract and vitamins from sterile anoxic stock solutions prepared under 100% N2 gas. Stock solutions of vitamins and fumarate should be sterilized by filtration. Adjust pH of complete medium to 7.0, if necessary.
962a THERMOVENABULUM MEDIUM