Medium data

Search for medium no. = [521].

Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

521 MODIFIED DESULFOGLAEBA ALKANEXEDENS MEDIUM

Mineral medium:

Sea salts (Sigma)	`25.0`	g
NH₄Cl	`0.25`	g
Na₂SO₄	`3.5`	g
Resazurin	`1.0`	mg
Distilled water	`900.0`	ml

Mix components and adjust pH to 7.2 - 7.4. Boil Mineral medium and cool down under a N₂-CO₂ (4:1, v/v) gas stream. Distribute Mineral medium into culture vessels (e.g., 9.0 ml of the medium in tubes) under the same gas mixture, seal with Teflon-coated rubber stoppers and autoclave. After cooling, add the following components (per 9.0 ml Mineral medium) from sterile anoxic stock solutions (autoclaved or *filter-sterilized):

8.5% NaHCO₃*	`0.4`	ml
2% Sodium pyruvate solution*	`0.5`	ml
Trace vitamins (see Medium No. 197)*	`0.1`	ml
Decane	`0.01`	ml
1% yeast extract solution	`0.05`	ml
Reducing agent solution (see below)	`0.08`	ml

Reducing agent solution:

Na₂S·9H₂O	`12.5`	g
L-Cysteine·HCl·H₂O	`12.5`	g
Distilled water	`1.0`	L

Autoclave and store under a N₂ atmosphere.
Comment: For strain JCM 13588, inoculate with a 20% inoculum, and incubate the vessels in inverted position.