435 ANAEROLINEA MEDIUM-B
Use Solution A of Medium No. 284 supplemented with final 1.0 ml Trace vitamins solution and 1.0 g yeast extract, and dissolved in 1.0 L distilled water. Dispense the medium into culture vessels (e.g., 20 ml of the medium in 50 ml serum bottle) under a stream of N2-CO2 (80:20, v/v) and seal with butyl rubber stoppers. After autoclaving, stand the medium overnight. Prior to inoculation, aseptically and anaerobically add 0.01 volume of L-cysteine·HCl·H2O and Na2S·9H2O solutions to the medium (3% solutions, autoclaved and stored under a N2 atmosphere).