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Main sol. 110a

Internal Solution-ID: S188

Equipment needed: Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
Casitone 30.0 g 29.703 -
Yeast extract 5.0 g 4.951 -
K2HPO4 5.0 g 4.951 28.422
Sodium resazurin
(0.1% w/v)
0.5 ml 5.0e-4 0.002
L-Cysteine HCl x H2O 0.5 g 0.495 2.819
Sugar mix 10.0 ml - -
Na2CO3 1.5 g 1.485 14.012
Meat filtrate 1000.0 ml - -
1 Dissolve ingredients, except cysteine, sugar mix, and carbonate, boil medium for 1 min, then cool to room temperature under 80% N2 and 20% CO2 gas atmosphere, add 0.5 g/l L-cysteine-HCl x H2O and dispense under same gas atmosphere into Hungate-type tubes (for strains demanding meat particles put those first into the tube, use 1 part meat particles to 4 or 5 parts fluid). Autoclave at 121°C for 20 min. After autoclaving add the sugar mix from a sterile anoxic stock solution prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Adjust the pH of the medium to pH 7.0, if necessary.
110a CMC MEDIUM (N2/CO2)