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.