# J825: KOKO MEDIUM


## Main sol. J825

1	g	Tryptone (BD-Difco)	
1	g	Peptone	
1	g	Yeast extract	
1.6	g	K2HPO4	
1	g	NaH2PO4 x 2 H2O	
0.5	g	NH4Cl	
0.16	g	MgSO4 x 7 H2O	
0.06	g	CaCl2 x 2 H2O	
10	ml	**Trace element solution SL-4**	
10	ml	**Trace vitamins**	
0.5	mg	Resazurin	
930	ml	Distilled water	

1. Mix components thoroughly and adjust pH to 7.0. Autoclave the medium under a N2 atmosphere. Add the following solutions from sterile anaerobic stocks (autoclaved or *filter-sterilized): 

50	ml	Glucose (10%)	
1.25	ml	NaHCO3 (*, 8%)	
10	ml	L-Cysteine HCl x H2O (3%)	
10	ml	Na2S x 9 H2O (3%)	


## Trace element solution SL-4

0.5	g	EDTA	
0.2	g	FeSO4 x 7 H2O	
100	ml	**Trace element solution SL-6**	
900	ml	Distilled water	


## Trace vitamins

2	mg	Biotin	
2	mg	Folic acid	
10	mg	Pyridoxine hydrochloride	
5	mg	Thiamine HCl	
5	mg	Riboflavin	
5	mg	Nicotinic acid	
5	mg	Calcium pantothenate	
0.1	mg	Vitamin B12	
5	mg	p-Aminobenzoic acid	
5	mg	Lipoic acid	
1000	ml	Distilled water	


## Trace element solution SL-6

0.1	g	ZnSO4 x 7 H2O	
0.03	g	MnCl2 x 4 H2O	
0.3	g	H3BO3	
0.2	g	CoCl2 x 6 H2O	
0.01	g	CuCl2 x 2 H2O	
0.02	g	NiCl2 x 6 H2O	
0.03	g	Na2MoO4 x 2 H2O	
1000	ml	Distilled water