Medium data

Search for medium no. = [378].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.


Yeast extract (BD-Difco)  2.0     g
Tryptone (BD-Difco)  2.0     g
Crotonic acid  0.86    g
KH2PO4  0.5     g
MgCl2·6H2O  0.33    g
NaCl  0.4     g
NH4Cl  0.4     g
CaCl2·2H2O  0.05    g
Trace metal solution (see below)  10.0     ml
Trace vitamins (see Medium No. 197)  10.0     ml
NaHCO3  3.5     g
L-Cysteine·HCL·H2O  0.5     g
Na2S·9H2O  0.5     g
Resazurin  1.0     mg
Dissolve ingredients, except crotonic acid, NaHCO3, L-cysteine·HCl·H2O and Na2S·9H2O, in 900 ml of distilled water and adjust pH to 7.0-7.5. 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 medium. Dispense 0.9 volume of the medium into suitable culture vessels under a stream of N2-CO2 (80:20, v/v) and seal with butyl rubber stoppers. After autoclaving, stand the medium overnight. Add 0.1 volume of a filter-sterilized crotonic acid solution (0.86 g/100 ml, pH 7.0) to the medium. Autoclave cysteine·HCl and Na2S·9H2O as 5% solutions, respectively, under a N2 atmosphere. Prior to inoculation, aseptically and anaerobically add the solutions of crotonic acid, L-cysteine·HCl·H2O and Na2S·9H2O to the medium.
Trace metal solution:
MnSO4·xH2O  1.0     g
Fe(NH4)2(SO4)2·6H2O  0.8     g
CoCl2·6H2O  0.2     g
ZnSO4·7H2O  0.2     g
CuCl2·2H2O  0.02    g
NiCl2·6H2O  0.02    g
Na2MoO4·2H2O  0.02    g
Na2SeO4  0.02    g
Na2WO4  0.02    g
Nitrilotriacetic acid  2.0     g
Distilled water  1.0     L
Adjust pH to 6.0 with KOH.