Medium data

Search for medium no. = [1146].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

1146   METHANIMICROCOCCUS MEDIUM

NH4Cl  0.5     g
MgSO4·7H2O  0.5     g
CaCl2·2H2O  0.25    g
NaCl  2.25    g
FeSO4·7H2O  0.002   g
FeCl2 solution(see Medium No. 187)  1.0     ml
Trace element solution(see Medium No. 187)  1.0     ml
Yeast extract (Oxoid)  2.0     g
Casitone (BD-BBL)  2.0     g
Sodium acetate  1.0     g
Resazurin  0.5     mg
Distilled water  925.0     ml
Mix components and autoclave under a N2-CO2 (80:20, v/v) gas atmosphere. After cooling, add the following solutions from anaerobic stocks (autoclaved or filter-sterilized):
Phosphate solution (see below)  10.0     ml
8% NaHCO3 solution*  35.0     ml
50% (v/v) Methanol  20.0     ml
1.4% Coenzyme M solution*  10.0     ml
Trace vitamins* (see Medium No. 197)  10.0     ml
Aseptically and anaerobically distribute the medium into culture vessels under a H2-CO2 (80:20, v/v) gas stream and seal with butyl stoppers. Prior to inoculation, add per liter the following solutions (autoclaved and stored under a N2 atmosphere):
5% Na2S·9H2O solution  7.2     ml
5% L-Cysteine·HCl·H2O solution  7.2     ml
Readjust pH to 7.0 - 7.2, if necessary. Pressurize inoculated culture vessels to 100 kPa H2-CO2 (80:20, v/v).
Phosphate solution:
K2HPO4  0.35    g
KH2PO4  0.23    g
Distilled water  10.0     ml



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