Trypticase Soy Agar………………………………..40.0 g
Hemin Solution (see below)………………………10.0 ml
DI Water……………………………………………..950.0 ml
Mix well and heat to boiling to dissolve.
Autoclave at 121?C. Cool to 50?C and aseptically add the following:
N-acetyl muramic acid (NAM) solution (see below)……..1.0 ml
Defibrinated Sheep Blood………………………………….50.0 ml
Aseptically dispense under a gas phase of 80% N2, 10% CO2, and 10% H2.
Hemin Solution
Hemin…………………………………50.0mg
K2HPO4………………………………1.74g
DI Water………………………………100.0 ml
Heat to boiling to dissolve.
N-acetyl muramic acid Solution
N-acetyl muramic acid……………………………….100.0 mg
DI Water………………………………………………..10.0 ml
Filter-sterilize. Store in small aliquots at -20 C.