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Main sol. 900

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Internal Solution-ID: S1852

Equipment needed: Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
KCl 0.33 g 0.329 4.418
MgCl2 x 6 H2O 0.52 g 0.519 2.553
CaCl2 x 2 H2O 0.30 g 0.299 2.037
NH4Cl 0.33 g 0.329 6.157
KH2PO4 0.33 g 0.329 2.42
Trace element solution SL-10 1.00 ml - -
Yeast extract 1.00 g 0.998 -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
Na2CO3 1.50 g 1.497 14.124
D-Fructose 5.00 g 4.99 27.698
Seven vitamins solution 1.00 ml - -
L-Cysteine HCl x H2O 0.30 g 0.299 1.705
Na2S x 9 H2O 0.30 g 0.299 1.247
Distilled water 1000.00 ml - -
1 Dissolve ingredients except carbonate, fructose, vitamins and reducing agents. Sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic, then dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add fructose, vitamins, cysteine 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 mixture. The pH of the complete medium should be at 7.0.
900 ACETOBACTERIUM TUNDRAE MEDIUM