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Main sol. 1310

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Internal Solution-ID: S2617

Equipment needed: Hungate tubes Autoclave Gassing station Centrifuge
Compound Amount Unit Conc. [g/L] Conc. [mM]
Casitone
(BD Bacto) 		10.00 g 10 -
Yeast extract
(BD Bacto) 		2.50 g 2.5 -
D-Glucose 2.00 g 2 11.101
Cellobiose 2.00 g 2 5.843
Maltose 2.00 g 2 5.843
Na-DL-lactate 5.00 g 5 44.619
Clarified rumen fluid 200.00 ml - -
K2HPO4 0.05 g 0.05 0.287
KH2PO4 0.05 g 0.05 0.367
(NH4)2SO4 0.10 g 0.1 0.757
NaCl 0.10 g 0.1 1.711
CaCl2 x 2 H2O 0.01 g 0.01 0.068
MgSO4 x 7 H2O 0.01 g 0.01 0.041
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 -
Na2CO3 4.00 g 4 37.74
L-Cysteine HCl x H2O 0.30 g 0.3 1.708
Na2S x 9 H2O 0.30 g 0.3 1.249
Distilled water 800.00 ml - -
1 Dissolve ingredients (except carbonate, cysteine and sulfide), boil medium for 1 min, then cool to room temperature under 100% CO2 gas atmosphere. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Add cysteine and sulfide form sterile anoxic stock solutions prepared under 100% N2 gas. Adjust pH of the complete medium to 6.5 - 6.8, if necessary.
1310 BUTYRIVIBRIO M2 MEDIUM