Medium data

Search for medium no. = [385].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

385   METHANOSARCINA ACETIVORANS MEDIUM

NaCl  23.4     g
MgSO4·7H2O  9.45    g
Na2CO3  5.0     g
NH4Cl  1.0     g
KCl  0.8     g
CaCl2·2H2O  0.14    g
Na2HPO4  0.6     g
Yeast extract (BD-Difco)  1.0     g
Trace minerals (see Medium No. 151)  10.0     ml
Methanol  6.5     ml
L-Cysteine·HCl·H2O  0.3     g
Na2S·9H2O  0.3     g
Resazurin  1.0     mg
Distilled water  1.0     L
Mix ingredients, except methanol, L-cysteine·HCl·H2O and Na2S·9H2O, and bring to a boil for 5-10 sec. Cool down under a stream of N2-CO2 (80:20), and add methanol to the medium, and adjust pH to 7.0 with HCl. Dispense the medium into suitable culture vessels (e.g., 20 ml of the medium in 120 ml serum bottles) under a stream of N2-CO2 (80:20) and seal with butyl rubber stoppers. After autoclaving, stand the medium overnight. Separately autoclave 5% L-cysteine·HCl·H2O and 5% Na2S·9H2O solutions under a N2 atmosphere. Prior to inoculation, aseptically and anaerobically add the L-cysteine·HCl·H2O and Na2S·9H2O solutions to the medium.