Medium data

Search for medium no. = [197].

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

197

KH₂PO₄	`0.75`	g
K₂HPO₄	`0.75`	g
MgCl₂·6H₂O	`0.36`	g
NH₄Cl	`1.0`	g
Yeast extract (BD-Difco)	`0.1`	g
Polypepton (Nihon Pharm. Co.)	`0.1`	g
Sodium acetate	`2.5`	g
Trimethylamine·HCl	`8.0`	g
Trace mineral solution (see below)	`9.0`	ml
Trace vitamins (see below)	`10.0`	ml
NaHCO₃	`0.8`	g
NaS₂·9H₂O	`0.5`	g
L-Cysteine·HCl·H₂O	`0.5`	g
Resazurin	`1.0`	mg
Distilled water	`981.0`	ml

Mix ingredients except NaHCO₃, Trace vitamins, Na₂S·9H₂O and cysteine·HCl·H₂O, and autoclave under a N₂-CO₂ (80:20) atmosphere. Filter-sterilize NaHCO₃ as an 8% solution and Trace vitamins while gassing with N₂. Separately autoclave Na₂S·9H₂O and cysteine·HCl·H₂O as 5% solutions under a N₂ atmosphere. Aseptically and anaerobically add NaHCO₃ and Trace vitamins to the medium under a N₂-CO₂ (80:20) atmosphere. Prior to inoculation, add the sterile Na₂S·9H₂O and cysteine·HCl·H₂O solutions, and adjust pH to 6.8.
Trace vitamins:

Biotin	`2.0`	mg
Folic acid	`2.0`	mg
Pyridoxine·HCl	`10.0`	mg
Thiamine·HCl	`5.0`	mg
Riboflavin	`5.0`	mg
Nicotinic acid	`5.0`	mg
Calcium pantothenate	`5.0`	mg
Vitamin B₁₂	`0.1`	mg
p-Aminobenzoic acid	`5.0`	mg
Lipoic acid	`5.0`	mg
Distilled water	`1.0`	L

Trace mineral solution:

Nitrilotriacetic acid	`4.5`	g
FeCl₂·4H₂O	`0.4`	g
MnCl₂·4H₂O	`0.1`	g
CoCl₂·6H₂O	`0.17`	g
ZnCl₂	`0.1`	g
CaCl₂·2H₂O	`0.02`	g
H₃BO₃	`0.019`	g
Na₂MoO₄·2H₂O	`0.01`	g
NiCl₂·6H₂O	`0.03`	g
Distilled water	`1.0`	L

Adjust pH to 7.0 with 1 N KOH.