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Main sol. 879

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Internal Solution-ID: S1820

Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 1.00 g 0.989 18.492
KCl 0.10 g 0.099 1.327
MgSO4 x 7 H2O 0.20 g 0.198 0.803
NaCl 0.80 g 0.791 13.54
KH2PO4 0.10 g 0.099 0.727
CaCl2 x 2 H2O 0.02 g 0.02 0.135
Yeast extract 1.00 g 0.989 -
Modified Wolin's mineral solution 10.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 4.9e-4 0.002
NaHCO3 1.00 g 0.989 11.774
D-Fructose 5.00 g 4.946 27.451
Wolin's vitamin solution (10x) 1.00 ml - -
L-Cysteine HCl x H2O 0.30 g 0.297 1.689
Na2S x 9 H2O 0.30 g 0.297 1.235
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except bicarbonate, fructose, vitamins and reducing agents), sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic and adjust pH to 5.5. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave at 121°C for 15 min. Add fructose, vitamins (sterilized by filtration), cysteine and sulfide from sterile stock solutions prepared under 100% N2 gas atmosphere and bicarbonate from a sterile stock solution prepared under 80% N2 and 20% CO2 gas atmosphere. Adjust pH of complete medium to 5.9!
879 CLOSTRIDIUM LJUNGDAHLII MEDIUM