ATCC Medium: 2751 Modified Chopped Meat Medium w/ 10mM Maltose
Chopped Meat Broth
Ground beef (fat-free).................500.0 g
Distilled water............................1000 ml
1N NaOH..................................25 ml
Mix meat, water and NaOH and bring to a boil with stirring. Simmer for at least 3 hours. Cool to room temperature, skim fat from surface, and filter. To filtrate, add sufficient distilled water to restore volume to 1.0 L.
To this filtrate, add:
Complete Medium
Trypticase Peptone (BD 211921)………....30.0 g
Yeast Extract................................................5.0 g
K2HPO4......................................................5.0 g
Maltose………………………………….……3.42 g
0.025% Resazurin.......................................4.0 ml
Agar (if necessary).....................................20.0 g
Boil and cool medium under 80% N 2 , 10% H2 , 10% CO2 , and add:
L-Cysteine HCl.........................................0.5 g
Hemin Solution (see below).......................10.0 ml
Vitamin K1 Solution (see below)...............0.2 ml
Adjust medium for final pH 7.0. Anaerobically dispense 7 ml into tubes.
Vitamin K1 Solution
Vitamin K1.........................0.15 ml 95% Ethanol.......................30.0 ml
Store solution in brown bottle under refrigeration. Discard after one month.
Hemin Solution
Hemin.............................50.0 mg
1N NaOH............................1.0 ml
Distilled water to...............100.0 ml
Autoclave complete medium at 121?C.