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Main sol. 78b

Internal Solution-ID: S129

Equipment needed: Filter Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
Ground beef
(fat free)
500.0 g 500 -
NaOH
(1 N)
25.0 ml - -
Casitone 30.0 g 30 -
Yeast extract 5.0 g 5 -
K2HPO4 5.0 g 5 28.707
Sodium resazurin
(0.1% w/v)
0.5 ml 5.0e-4 0.002
L-Cysteine HCl x H2O 0.5 g 0.5 2.847
Distilled water 1000.0 ml - -
1 Use lean beef or horse meat. Remove fat and connective tissue before grinding. Mix meat, water and NaOH, then boil for 15 min with stirring. Cool to room temperature, skim fat off surface, and filter, retaining both meat particles and filtrate. To the filtrate add Casitone, yeast extract, hydrogen phosphate and resazurin, then add water to a final volume of 700 ml. Sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic, then add cysteine and adjust pH to 7.0. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes containing meat particles (use ca. 7 ml liquid medium for 1 - 2 g meat particles), then autoclave at 121°C for 20 min.
2 In some cases (as indicated in the catalogue) the addition of Haemin and Vitamin K1 or Vitamin K3 is necessary. Add to 1000 ml of medium after autoclaving:
Haemin solution
optional
10.0 ml - -
Vitamin K1 solution
optional
10.0 ml - -
Vitamin K3 solution
optional
1.0 ml - -
78b CHOPPED MEAT MEDIUM (N2/CO2)