ATCC medium: 639 Heart infusion agar or broth, pH 7.5, containing 20% (v/v) inactivated- and acid-adjusted porcine serum and 10% (v/v) yeast extract
Prepare Heart Infusion Agar (BD 244400) or Heart Infusion Broth (BD 238400) according to manufacturer’s instructions.
Add 20% (v/v) inactivated- and acid-adjusted porcine serum (see below) and 10% (v/v) Yeast Extract (GIBCO 360-8180).
Acid Adjusted Porcine Serum:
Adjust pH of Porcine Serum (GIBCO 26250) to 4.4 with sterile 1 N HCl taking care not to drop pH below 4.2.
Allow serum to stand 18-20 hours at 4C. Adjust serum to pH 7.0 with sterile 1 N NaOH. Centrifuge at 9,000 rpm for 20 minutes and discard sediment.
Filter through #02 Selas (0.85 millimicrons pore size) for clearing. Filter-sterilize. Store at -70C until ready for use.