Medium data

Search for medium no. = [1103].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.


Solution A:
KH2PO4  0.14    g
MgCl2·6H2O  0.2     g
CaCl2·2H2O  0.15    g
NH4Cl  0.54    g
Yeast extract (BD-Difco)  2.3     g
Trace element solution SL-11 (see below)  1.0     ml
Selenite tungstate solution (see Medium No. 431)  0.5     ml
Trace vitamins (see Medium No. 197)  10.0     ml
NaHCO3  2.5     g
Resazurin  1.0     mg
Distilled water  900.0     ml
Mix components except NaHCO3 and trace vitamins, and adjust pH to 6.5. Bring to a boil for 5-10 sec, then, cool down under a stream of N2-CO2 (80:20, v/v) and add NaHCO3 to the solution. Dispense Solution A into culture vessels under a stream of N2-CO2 (80:20, v/v), seal with butyl rubber stopper and autoclave. After autoclaving, stand the medium overnight. Add trace vitamins from a sterile anaerobic stock (filter-sterilized).
Solution B:
Glucose  2.2     g
Distilled water  100.0     ml
Adjust pH to 7.0 and filter-sterilize. Keep under a N2 atmosphere.
To complete the medium, aseptically and anaerobically add 0.1 volume of Solution B to 0.9 volume of Solution A (e.g., 18 ml Solution A in 60 ml serum bottles and 2 ml Solution B). Prior to inoculation, add per liter the following solutions:
5% Na2S·9H2O solution  6.0     ml
5% L-Cysteine·HCl·H2O solution  6.0     ml
Trace element solution SL-11:
EDTA·2Na  5.2     g
FeCl2·4H2O  1.5     g
ZnCl2  70.0     mg
MnCl2·4H2O  100.0     mg
H3BO3  6.0     g
CoCl2·6H2O  190.0     mg
CuCl2·2H2O  2.0     mg
NiCl2·6H2O  24.0     mg
Na2MoO4·2H2O  36.0     mg
Distilled water  1.0     L
Adjust pH to 6.0.

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