Heart Infusion Broth………………………………………17.5 g
Agar Noble (if required)…………………...………………10.0 g
DI Water…………………………………………………….700 ml
Autoclave at 121?C. Cool to 55C and aseptically add the following:
Horse serum (heat-inactivated)………………………….200.0 ml
Yeast Extract #3 (ATCC 9678, see below)… …..……..100.0 ml
**Horse serum should be heat inactivated at 56?C for 30 minutes. When making with agar, be sure to warm serum and yeast extract solution to 55?C before adding to basal medium.
Final pH should be 7.4. Dispense as desired.
Yeast (Store bought Fleischmann’s yeast)…….………….150 g
Yeast Extract #3
DI Water……………………………………………………….1000 ml
Mix together for 10 minutes, autoclave at 121?C. Let stand overnight. Transfer yeast solution to centrifuge bottles and spin at 9,000 rpms for 50 minutes. Carefully pour supernatant into 100 ml bottles and discard the cell debris. Autoclave at 121?C and store sterile at 2-8?C.
*Note: As of 10/26/2011 Gibco Yeast Extract is no longer in use. Gibco changed the supplier of their raw materials as of early 2011. This new version no longer supports growth and has been replaced with Yeast Extract #3.