Medium data

Search for medium no. = [492].

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

492 MMJ-TL MEDIUM

Modified MJ synthetic seawater (see below)	`1.0`	L
Sulfur (powder)	`3.0`	g
Resazurin	`1.0`	mg

Mix ingredients except sulfur, adjust pH to 7.0 with NaOH, and autoclave under a N₂ atmosphere. After cooling, add the following solutions (autoclaved or *filter-sterilized):

Trace vitamins* (see Medium No. 197)	`10.0`	ml
1.0 M Sodium acetate solution	`10.0`	ml
1% Yeast extract solution	`10.0`	ml

Steam sulfur for 3 hr on each of 3 successive days. Distribute the medium into culture vessels (e.g. 20 ml in 120 ml serum bottles) containing sulfur under a stream of a H₂-CO₂ (4:1, v/v) gas mixture, and seal with butyl rubber stoppers. Prior to inoculation, add per liter the following solution (autoclaved and stocked under a N₂ atmosphere):

5% Na₂S·9H₂O solution 10.0 ml

Pressurize the inoculated bottles to 200 kPa H₂-CO₂ (4:1, v/v).
Modified MJ synthetic seawater:

NaCl	`30.0`	g
K₂HPO₄	`0.14`	g
CaCl₂·2H₂O	`0.5`	g
NH₄Cl	`0.5`	g
MgSO₄·7H₂O	`3.4`	g
MgCl₂·6H₂O	`4.18`	g
KCl	`0.33`	g
FeSO₄·7H₂O	`0.01`	g
NiCl₂·6H₂O	`1.0`	mg
Na₂SeO₃·5H₂O	`1.0`	mg
Trace minerals (see Medium No. 151)	`10.0`	ml
Distilled water	`1.0`	L