Medium data

Search for medium no. = [1331].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

1331   HYDROGENOTROPHIC METHANOGEN MEDIUM

NH4Cl  0.54    g
KH2PO4  0.14    g
MgCl2·6H2O  3.0     g
CaCl2·2H2O  0.15    g
NaCl  20.0     g
FeCl2 solution (see Medium No. 187)  1.0     ml
Trace element solution (see Medium No. 187)  1.0     ml
Selenite-tungstate solution (see Medium No. 431)  1.0     ml
Sodium acetate·3H2O  0.27    g
NaHCO3  2.5     g
Resazurin  1.0     mg
Distilled water  1.0     L
Mix components, except NaHCO3, and adjust pH 7.0. Bring to a boil for 5-10 seconds, 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., 20 ml in 120 ml serum bottles) under a stream of H2-CO2 (80:20 v/v), seal with butyl rubber stoppers and autoclave. Aseptically and anaerobically add per liter the following solutions (autoclaved or filter-sterilized, and stored under a N2 atmosphere):
Trace vitamins* (see Medium No. 197)  2.0     ml
0.5 M Coenzyme M solution  10.0     ml
Titanium citrate solution* (see below)  10.0     ml
Titanium citrate solution:
Titanium(III) chloride  0.75    g
Sodium citrate·2H2O  2.94    g
Distilled water  55.0     ml
Mix components and centrifuge at 8,000 xg for 10 min. Filter-sterilize the supernatant and store under a N2 atmosphere.



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