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Main sol. 860

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Internal Solution-ID: S1759

Equipment needed: Hungate tubes Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 1.0 g 0.998 18.658
K2HPO4 0.3 g 0.299 1.719
KH2PO4 0.3 g 0.299 2.2
MgCl2 x 6 H2O 0.2 g 0.2 0.982
CaCl2 x 2 H2O 0.1 g 0.1 0.679
KCl 0.1 g 0.1 1.339
NaCl 0.6 g 0.599 10.246
Yeast extract 1.0 g 0.998 -
Trace element solution SL-10 1.5 ml - -
Sodium resazurin
(0.1% w/v) 		0.5 ml 5.0e-4 0.002
Na2CO3 1.5 g 1.497 14.124
D-Fructose 4.0 g 3.992 22.158
L-Cysteine HCl x H2O 0.5 g 0.499 2.841
Na2S x 9 H2O 0.3 g 0.299 1.247
Distilled water 1000.0 ml - -
1 Dissolve ingredients except carbonate, fructose, cysteine and sulfide, then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add fructose, 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. Prior to use adjust pH of complete medium to 7.0 - 7.2, if necessary.
860 ANAEROCOLUMNA MEDIUM