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Main sol. 1175

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Internal Solution-ID: S2347

Equipment needed: Autoclave Hungate tubes Anaerobic workbench
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 0.20 g 0.2 3.739
MgCl2 x 6 H2O 0.05 g 0.05 0.246
KH2PO4 0.20 g 0.2 1.47
Na2CO3 7.40 g 7.4 69.819
NaHCO3 18.50 g 18.5 220.22
Na2S x 9 H2O 0.50 g 0.5 2.082
Yeast extract 0.20 g 0.2 -
Cellobiose 3.00 g 3 8.764
Distilled water 1000.00 ml - -
1 pH 10.0
2 Prepare the medium without Na2CO3, NaHCO3, Na2S x 9 H2O, cellulobiose or yeast extract. Boil the medium in a water bath and cool under a stream of nitrogen. Add the Na2CO3 and NaHCO3 to the cooled medium (the pH should be about 10.0) and dispense the medium into Hungate tubes or serum bottles under a stream of nitrogen. Seal the tubes under nitrogen gas and autoclave. To the cooled medium at the appropriate amounts of Na2S x 9 H2O, cellulobiose and yeast extract from sterile, anaerobic stock solutions (10% Na2S x 9 H2O, 5% cellulobiose, 10% yeast extract).
1175 ALKALIFLEXUS MEDIUM