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Main sol. 902

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Internal Solution-ID: S1855

Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 20.00 g 19.96 341.548
KCl 0.70 g 0.699 9.371
MgSO4 x 7 H2O 3.90 g 3.892 15.792
CaCl2 x 2 H2O 0.40 g 0.399 2.715
NH4Cl 0.30 g 0.299 5.597
Na2HPO4 0.15 g 0.15 1.055
Na2SiO3 0.03 g 0.03 0.245
Yeast extract 0.05 g 0.05 -
Trace element solution SL-10 1.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
NaHCO3 0.50 g 0.499 5.94
Na-pyruvate 2.50 g 2.495 22.673
Wolin's vitamin solution (10x) 1.00 ml - -
L-Cysteine HCl x H2O 0.45 g 0.449 2.557
Na2S x 9 H2O 0.45 g 0.449 1.87
Distilled water 1000.00 ml - -
1 Dissolve ingredients except bicarbonate, pyruvate, vitamins and reducing agents, then adjust pH of solution to 7.0 with NaOH. Sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic, then dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add pyruvate, vitamins, cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Vitamins should be sterilized by filtration. Adjust pH of complete medium to 7.0, if necessary.
902 CARCBOXYDIBRACHIUM MEDIUM