# J967: KD-1 MEDIUM


## Main sol. J967

10	g	Bacto peptone (BD-Difco)	
2	g	Urea	
2.5	g	K2HPO4	
3	g	KH2PO4	
3	g	NaCl	
0.2	g	MgSO4 x 7 H2O	
10	ml	**Trace vitamins**	
10	ml	**Trace element solution**	
1	mg	Resazurin	
1000	ml	Distilled water	

1. Mix components thoroughly and adjust pH to 8.0. Distribute the medium into culture vessels under a N2-CO2 (9:1, v/v) gas mixture, seal with butyl rubber stoppers and autoclave.


## 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

1.5	g	Nitrilotriacetic acid	
3	g	MgSO4 x 7 H2O	
0.5	g	MnSO4 x n H2O	
1	g	NaCl	
0.1	g	FeSO4 x 7 H2O	
0.18	g	CoSO4 x 7 H2O	
0.1	g	CaCl2 x 2 H2O	
0.18	g	ZnSO4 x 7 H2O	
10	mg	CuSO4 x 5 H2O	
20	mg	AlK(SO4)2 x 12 H2O	
10	mg	H3BO3	
10	mg	Na2MoO4 x 2 H2O	
25	mg	NiCl2 x 6 H2O	
0.3	mg	Na2SeO3 x 5 H2O	
1000	ml	Distilled water	

1. Dissolve nitrilotriacetic acid at first and adjust pH to 6.5 with KOH, then add the remaining minerals.
2. Adjust final pH of the medium to 7.0 with KOH.