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Main sol. 516

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Internal Solution-ID: S1143

Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 0.33 g 0.329 6.157
KH2PO4 0.33 g 0.329 2.42
KCl 0.33 g 0.329 4.418
MgCl2 x 6 H2O 0.33 g 0.329 1.62
CaCl2 x 2 H2O 0.33 g 0.329 2.24
Trace element solution SL-10 1.00 ml - -
Yeast extract 0.50 g 0.499 -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
Na2CO3 1.50 g 1.497 14.124
Cellobiose 5.00 g 4.99 14.578
Wolin's vitamin solution (10x) 1.00 ml - -
Na2S x 9 H2O 0.50 g 0.499 2.078
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except vitamins, carbonate, cellobiose and sulfide), 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. Prior to inoculation add vitamins, cellobiose 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. Stock solutions of vitamins and cellobiose should be sterilized by filtration. The pH of the complete medium should be at 7.1 - 7.3.
516 ANAEROCELLUM MEDIUM