ATCC medium: 1482 PYG plus 0.1% Tween 80
Tween 80.................................1.0 g
Glucose.................................10.0 g
Peptone (BD 211677)......................5.0 g
Trypticase Peptone (BD 211921)...........5.0 g
Yeast extract...........................10.0 g
0.025% Resazurin.........................4.0 ml
Hemin Solution (see below)..............10.0 ml
Salts Solution (see below)............. 40.0 ml
Vitamin K1 Solution (see below)..........0.2 ml
L-Cysteine . HCl.........................0.5 g
Distilled water........................950.0 ml
Bring medium without cysteine to a boil; add cysteine; adjust medium for a final pH of 7.0; autoclave at 121C for 15 minutes.
Hemin Solution:
Hemin.............................50.0 mg
N NaOH.............................1.0 ml
Distilled water to...............100.0 ml
Dissolve hemin in NaOH. Add distilled water. Autoclave at 121C for 15 minutes.
Salts Solution:
CaCl2 ..............................0.2 g
MgSO4 ..............................0.2 g
K2HPO4 .............................1.0 g
KH2PO4 .............................1.0 g
NaHCO3 ............................10.0 g
NaCl...............................2.0 g
Distilled water....................1.0 L
Vitamin K1 Solution:
Vitamin K1.........................0.15 ml
95% Ethanol.......................30.0 ml
Store K1 solution in a brown bottle under refrigeration.
Discard solution after 1 month.