# J1278: MIZ13 MEDIUM


## Main sol. J1278

1000	ml	**Modified Wolfe's mineral solution**	
10	ml	**Trace minerals**	
1.95	g	MES	
0.83	g	NaHCO3	
1.36	g	Sodium acetate	
1.1	g	Sodium pyruvate	
3.2	g	Sodium fumarate	

1. Mix components thoroughly and adjust pH 6.2. Autoclave under a N2 atmosphere. After cooling, add the following solutions anaerobically and aseptically (autoclaved or *filter-sterilized and stored under a N2 atmosphere): 

10	ml	**Trace vitamins** (*)	
10	ml	L-Cysteine HCl x H2O (5%)	


## Modified Wolfe's mineral solution

1	g	NH4Cl	
0.2	g	MgSO4 x 7 H2O	
0.1	g	CaCl2 x 2 H2O	
0.05	g	KH2PO4	
1000	ml	Distilled water	


## Trace minerals

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	
0.1	g	CoSO4 x 7 H2O	
0.1	g	CaCl2 x 2 H2O	
0.1	g	ZnSO4 x 7 H2O	
0.01	g	CuSO4 x 5 H2O	
0.01	g	AlK(SO4)2	
0.01	g	H3BO3	
0.01	g	Na2MoO4 x 2 H2O	
1000	ml	Distilled water	

1. Dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH solution. Then proceed to add minerals. Adjust final pH to 7.0.


## 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