Medium data

Search for medium no. = [232].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

232   METHANOCALDOCOCCUS JANNASCHII MEDIUM

K2HPO4  0.14    g
CaCl2·2H2O  0.14    g
NH4Cl  0.25    g
MgSO4·7H2O  3.4     g
MgCl2·6H2O  2.7     g
KCl  0.33    g
NiCl2·6H2O  0.75    mg
Na2SeO3·5H2O  0.5     mg
NaCl  30.0     g
Fe(NH4)2(SO4)2·6H2O  0.01    g
Trace minerals (see Medium No. 151)  10.0     ml
Trace vitamins (see Medium No. 197)  10.0     ml
NaHCO3  1.0     g
L-Cysteine·HCl·H2O  0.5     g
Na2S·9H2O  0.5     g
Resazurin  1.0     mg
Distilled water  980.0     ml
Mix ingredients, except NaHCO3, cysteine·HCl and Na2S·9H2O, and adjust pH to 6.0. Bring to a boil for 5-10 sec, then, cool down under a stream of H2-CO2 (80:20) and add NaHCO3 to the medium. Dispense the medium into suitable culture vessels (e.g., 20 ml of the medium in 120 ml serum bottles) under a stream of H2-CO2 (80:20) and seal with butyl rubber stoppers. After autoclaving, stand the medium overnight. Separately autoclave cysteine·HCl and Na2S·9H2O as 5% solutions, respectively, under a N2 atmosphere. Prior to inoculation, aseptically and anaerobically add the cysteine·HCl and Na2S·9H2O solutions. Pressurize the inoculated bottles to 200 kPa H2-CO2 (80:20).