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|Ground beef (fat free)||500.0||g|
|NaOH 1 N||25.0||ml|
|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 water to a final volume of 1000 ml, and then add:|
|Boil, cool under nitrogen atmosphere, add 0.5 g/l cysteine and adjust pH to 7.0. To make medium anoxic boil it, cool under nitrogen atmosphere, add 0.5 g/l cysteine hydrochloride and adjust pH to 7.0. Dispense anoxically 7 ml medium into Hungate 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 30 min. For agar slants use 15 g agar per 1000.0 ml medium.|
|Haemin solution (see below)||10.0||ml|
|Vitamin K1 or Vitamin K3 solution (see below)||0.20||ml|
** ph : 7.0.
** memo : -