# J854: COLON EXTRACT MEDIUM


## Main sol. J854

167	ml	**Solution A**	
167	ml	**Solution B**	
40	ml	**Solution C**	
200	ml	**Solution D**	
5	g	Yeast extract	
5	g	Trypticase peptone	
5	g	NaHCO3	
1	mg	Resazurin	
1	g	L-Cysteine HCl x H2O	
0.2	ml	Vitamin K1 (0.5%)	
10	ml	**Hemin solution**	
1000	ml	Distilled water	

1. Adjust pH to 7.0.


## Solution A

5.4	g	NaCl	
2.7	g	KH2PO4	
0.16	g	CaCl2 x 2 H2O	
0.12	g	MgCl2 x 6 H2O	
0.07	g	MnSO4 x n H2O	
0.06	g	CoCl2 x 6 H2O	
5.4	g	(NH4)2SO4	
0.05	g	FeSO4 x 7 H2O	
1000	ml	Distilled water	


## Solution B

2.7	g	K2HPO4	
1000	ml	Distilled water	


## Solution C

50	g	Glucose	
50	g	Maltose	
50	g	Cellobiose	
50	g	Xylose	
1000	ml	Distilled water	


## Solution D

1. Dilute (1:1, w/v) the content of terminal pig colon with the following buffer: 167 ml Solution A (see above) and 167 ml Solution B (see above). Make up to 1.0 L with distilled water. Stomach the suspension and filter through a cheese cloth. Incubate at 37C for 48 h and adjust pH to 7.0. Centrifuge at 20,000 x g for 20 min. Retrieve the supernatant and store at -20C until use.


## Hemin solution

1. Dissolve 50 mg hemin first in 1 ml of 1 N NaOH. Then adjust volume to 100 ml.