Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.1130 YCFA MEDIUM
Tryptone (BD-Difco) 10.0 g
Yeast extract (BD-Difco) 2.5 g
NaHCO3 4.0 g
Glucose 2.0 g
Maltose 2.0 g
Cellobiose 2.0 g
L-Cysteine·HCl·H2O 1.0 g
Mineral solution I (see below) 150.0 ml
Mineral solution II (see below) 150.0 ml
Resazurin 1.0 mg
VFA mix (see below) 6.2 ml
Hemin solution (see below) 10.0 ml
Vitamin solution I (see below) 1.0 ml
Distilled water 660.0 ml
Adjust pH to 7.45.
After autoclaving, add filter-sterilized 1.0 ml Vitamin solution II (see below).
Mineral solution I:
K2HPO4 3.0 g
Distilled water 1.0 L
Mineral solution II:
KH2PO4 3.0 g
(NH4)2SO4 6.0 g
NaCl 6.0 g
MgSO4·7H2O 0.6 g
CaCl2·2H2O 0.6 g
Distilled water 1.0 L
VFA mix:
Acetic acid 17.0 ml
Propionic acid 6.0 ml
n-Valeric acid 1.0 ml
iso-Valeric acid 1.0 ml
iso-Butyric acid 1.0 ml
10 N NaOH 26.0 ml
Hemin solution:
KOH 0.28 g
Ethanol 25.0 ml
Hemin 0.1 g
Adjust volume to 100 ml with distilled water.
Vitamin solution I:
Biotin 5.0 mg
Vitamin B12 5.0 mg
p-Aminobenzoic acid 15.0 mg
Folic acid 25.0 mg
Pyridoxine·HCl 75.0 mg
Distilled water 500.0 ml
Vitamin solution II:
Thiamine·HCl 25.0 mg
Riboflavin 25.0 mg
Distilled water 500.0 ml