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Main sol. 960

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Internal Solution-ID: S1981

Equipment needed: Filter Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.14 g 0.14 1.027
MgCl2 x 6 H2O 0.20 g 0.2 0.982
CaCl2 x 2 H2O 0.15 g 0.15 1.018
NH4Cl 0.54 g 0.539 10.075
Trace element solution 1.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
Na2CO3 1.50 g 1.497 14.124
Na-pyruvate 2.20 g 2.196 19.952
Yeast extract
(OXOID) 		0.10 g 0.1 -
Wolin's vitamin solution (10x) 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, pyruvate, yeast extract, vitamins, and reducing agents) 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. Add pyruvate, yeast extract, 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 atmosphere. Stock solutions of vitamins and pyruvate should be sterilized by filtration. The pH of the complete medium should be 7.0.
2 Note: Use 10% (v/v) as inoculum.
960 PELOTOMACULUM MEDIUM