Medium data

Search for medium no. = [230].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

230   METHANOSARCINA MEDIUM

KH2PO4  0.227   g
K2HPO4  0.348   g
NH4Cl  0.5     g
MgSO4·7H2O  0.5     g
CaCl2·2H2O  0.25    g
NaCl  2.25    g
FeSO4·7H2O  2.0     mg
Yeast extract (BD-Difco)  2.0     g
Casitone (BD-Difco)  2.0     g
Trace vitamins (see Medium No. 197)  10.0     ml
FeCl2 solution (see Medium No. 187)  1.0     ml
Trace element solution (see Medium No. 187)  1.0     ml
NaHCO3  0.85    g
Methanol  10.0     ml
L-Cysteine·HCl·H2O  0.3     g
Na2S·9H2O  0.3     g
Resazurin  1.0     mg
Distilled water  980.0     ml
Mix ingredients, except NaHCO3, methanol, cysteine·HCl and Na2S·9H2O, and bring to a boil for 5-10 sec. Cool down under a stream of N2-CO2 (80:20), and add NaHCO3 and methanol 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 N2-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.