Medium data

Search for medium no. = [412].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

412   Fe(III)-REDUCER FRESHWATER ENRICHMENT MEDIUM

Poorly crystalline iron(III) oxide (see Medium No. 411)  100.0     mM
MgCl2·6H2O  0.33    g
CaCl2·2H2O  0.33    g
KCl  0.33    g
KH2PO4  0.33    g
NH4Cl  0.25    g
NaHCO3  2.5     g
Na2SeO4  18.9     mg
Trace vitamins (see Medium No. 197)  10.0     ml
Trace mineral solution (see Medium No. 411)  10.0     ml
L-Cysteine·HCl·H2O  0.044   g
FeCl2·4H2O  0.26    g
Add components except L-cysteine·HCl·H2O and FeCl2·4H2O to distilled water, bring volume to 1.0 L and adjust pH to 7.0. Distribute the medium into appropriate culture vessels (e.g., 20 ml of the medium in 120 ml serum bottles) under a H2-CO2 (4:1, v/v) gas mixture, sparge with the same gas mixture for 10 - 15 min, seal with butyl rubber stoppers and autoclave. Separately autoclave concentrated (100- or 200-fold) L-cysteine·HCl·H2O and FeCl2·4H2O solutions under a N2 gas atmosphere. Prior to inoculation, add the sterile L-cysteine·HCl·H2O and FeCl2·4H2O solutions. Pressurize the inoculated vessels to 100 kPa H2-CO2 (4:1, v/v) and cultivate without shaking.
Comment: For strain JCM 12379, inoculate with a 10% inoculum.