Medium data

Search for medium no. = [1126].

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

1126 SULFATE-FREE METAKO MEDIUM

MgCl₂·6H₂O	`9.47`	g
CaCl₂·2H₂O	`1.36`	g
NaCl	`20.75`	g
KCl	`0.60`	g
NH₄Cl	`0.54`	g
KH₂PO₄	`0.14`	g
NaHCO₃	`2.69`	g
KBr	`85.0`	mg
Na₂B₄O₇·10H₂O	`34.0`	mg
SrCl₂·6H₂O	`12.0`	mg
NaF	`3.0`	mg
LiCl	`1.0`	mg
Trace element solution (see below)	`1.0`	ml
Selenite-tungstate solution (see Medium No. 431)	`0.4`	ml
Trace vitamins solution (see Medium No. 682)	`2.0`	ml
Resazurin	`1.0`	mg
Distilled water	`1.0`	L

Mix components thoroughly and distribute the medium into culture vessels (e.g., 20 ml medium in 100 ml serum bottles or 5.0 ml medium in Hungate tubes) under a stream of N₂-CO₂ (80:20, v/v), seal with butyl rubber stoppers and autoclave. Allow to stand the autoclaved medium overnight. Add per liter the following solution from a sterile anaerobic stock (filter-sterilized and stored under a N₂ atmosphere):

1 M Trimethylamine solution 10.0 ml

Prior to inoculation, add per liter the following solutions aseptically and anaerobically (autoclaved and stored under a N₂ atmosphere):

5% L-Cysteine·HCl·H₂O solution	`2.5`	ml
5% Na₂S·9H₂O solution	`2.5`	ml

Trace element solution:

Nitrilotriacetic acid	`0.15`	g
MnCl₂·4H₂O	`0.61`	g
CoCl₂·6H₂O	`0.42`	g
ZnCl₂	`0.09`	g
CuCl₂·2H₂O	`0.007`	g
AlCl₃	`0.006`	g
H₃BO₃	`0.01`	g
Na₂MoO₄·2H₂O	`0.02`	g
SrCl₂·6H₂O	`0.01`	g
NaBr	`0.01`	g
KI	`0.07`	g
FeCl₃·6H₂O	`0.5`	g
NiCl₂·6H₂O	`0.025`	g
Distilled water	`1.0`	L

Comment: For strain JCM 32479, replace trimethylamine with sodium lactate (final 0.55 g/L) and yeast extract (1.0 g/L).