Medium data

Search for medium no. = [284].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.


Solution A:
Yeast extract (BD-Difco)  0.1     g
KH2PO4  0.14    g
MgCl2·6H2O  0.2     g
CaCl2·2H2O  0.15    g
NH4Cl  0.54    g
Trace vitamins solution (see below)  2.0     ml
Trace element solution (see below)  1.0     ml
Se/W solution (see below)  1.0     ml
NaHCO3  2.5     g
Resazurin  1.0     mg
Distilled water  900.0     ml
Mix components except NaHCO3 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 0.9 volume of the medium into culture vessels (e.g., 18 ml of the medium in 50 ml serum bottle) under a stream of N2-CO2 (80:20, v/v) and seal with butyl rubber stoppers. After autoclaving, stand the medium overnight.
Solution B:
Sodium pyruvate  2.2     g
Distilled water  100.0     ml
Adjust pH to 7.0 and filter-sterilize. Keep the solution under a N2 atmosphere.
To complete the medium, add 0.1 volume of Solution B to the culture vessels containing Solution A. Prior to inoculation, aseptically and anaerobically add 0.01 volume of L-cysteine·HCl·H2O and Na2S·9H2O solutions to the medium (3% solutions, autoclaved and stored under a N2 atmosphere).
Comment: For strain JCM 10971, incubate with a 10% inoculum.
Trace vitamins solution:
Biotin  4.9     mg
Folic acid  8.8     mg
Pyridoxine·HCl  4.1     mg
Thiamine·HCl  6.7     mg
Riboflavin  7.5     mg
Nicotinic acid  2.5     mg
DL-Calcium pantothenate  9.5     mg
Vitamin B12  27.1     mg
p-Aminobenzoic acid  2.7     mg
Lipoic acid  4.1     mg
Distilled water  1.0     L
Trace element solution:
Nitrilotriacetic acid  12.8     g
FeCl3·6H2O  1.35    g
MnCl2·4H2O  0.1     g
CoCl2·6H2O  0.024   g
CaCl2·2H2O  0.1     g
ZnCl2  0.1     g
CuCl2·2H2O  0.025   g
H3BO3  0.01    g
Na2MoO4·2H2O  0.024   g
NiCl2·6H2O  0.12    g
Distilled water  1.0     L
Dissolve nitrilotriacetic acid first in 200 ml of distilled water and adjust pH to 6.5 with KOH. Then add the remaining salts and bring volume to 1.0 L. Readjust pH to 7.0 with KOH.
Se/W solution:
Na2SeO3·5H2O  4.0     mg
Na2WO4·2H2O  4.0     mg
Distilled water  1.0     L

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