# J779: MODIFIED HALOANAEROBIUM MEDIUM


## Main sol. J779

75	g	NaCl	
8.8	g	MgSO4 x 7 H2O	
1	g	KCl	
10	g	Yeast extract	
10	g	Trypticase peptone (BD-BBL)	
10	ml	**Trace vitamins**	
10	ml	**Trace minerals**	

1. Add components to distilled water and bring volume to 1.0 L. Adjust pH 8.0 with NaOH. Autoclave under a N2 atmosphere. After cooling, add anaerobically the following solutions (autoclaved or *filter-sterilized): 

25	ml	Glucose (10%)	
25	ml	**Thioglycolate-ascorbate solution** (*)	

2. Readjust pH to 8.0, if necessary.


## Thioglycolate-ascorbate solution

0.5	g	Sodium thioglycolate	
0.5	g	Sodium ascorbate	
25	ml	Distilled water	

1. Prepare just before use. Filter-sterilize.


## 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 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.