Medium data

Search for medium no. = [807].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

807   WR MEDIUM FOR METHANOBACTERIACEAE

KH2PO4  0.136   g
NH4Cl  0.535   g
MgCl2·6H2O  0.204   g
CaCl2·2H2O  0.147   g
NaHCO3  2.52    g
Tryptone (BD-Difco)  1.0     g
Sodium acetate  0.82    g
Trace element solution (see Medium No. 243)  10.0     ml
Trace vitamins (see Medium No. 197)  10.0     ml
Se/W solution (see Medium No. 284)  1.0     ml
L-Cysteine·HCl·H2O  0.12    g
Na2S·9H2O  0.12    g
Resazurin  1.0     mg
Distilled water  1.0     L
Mix ingredients except NaHCO3, L-cysteine·HCl·H2O and Na2S·9H2O, and adjust pH to 7.0. Bring to a boil for 5-10 sec, then, cool down under a stream of H2-CO2 (80:20, v/v) and add NaHCO3. Dispense the medium into suitable culture vessels (e.g., 5 ml in Hungate tubes) under the same gas mixture and seal with butyl rubber stoppers. After autoclaving, stand the medium overnight. Separately autoclave L-cysteine·HCl·H2O and Na2S·9H2O as 5% solutions, respectively, under a N2 atmosphere. Prior to inoculation, aseptically and anaerobically add L-cysteine·HCl·H2O and Na2S·9H2O solutions. Readjust pH to 6.7, if necessary. Pressurize the inoculated bottles to 150 kPa H2-CO2 (80:20, v/v).