# J368: MJAIS MEDIUM


## Main sol. J368

30	g	NaCl	
0.9	g	K2HPO4	
0.7	g	KH2PO4	
1	g	CaCl2 x 2 H2O	
1.25	g	NH4Cl	
3.4	g	MgSO4 x 7 H2O	
4.18	g	MgCl2 x 6 H2O	
0.33	g	KCl	
5	mg	NiCl2 x 6 H2O	
0.5	mg	Na2SeO3 x 5 H2O	
0.01	mg	Fe2(SO4)3 x n H2O	
5	mg	H2WO4	
0.02	g	FeSO4 x 7 H2O	
0.02	g	Ferric citrate	
10	ml	**Trace mineral solution**	
1	ml	**Trace vitamins**	
1	g	NaHCO3	
10	g	Sulfur (powder)	
0.5	g	Na2S x 9 H2O	
1	mg	Resazurin	
1000	ml	Distilled water	

1. Mix ingredients, except Trace vitamins, NaHCO3, sulfur and Na2S x 9H2O, adjust pH to 5.5, and autoclave. Filter-sterilize Trace vitamins and 5% NaHCO3 solution and add to the medium. Steam sulfur for 3 hr on each of 3 successive days. Separately autoclave 5% Na2S x 9H2O solution under a N2 atmosphere. Distribute the medium into culture vessels containing appropriate amount of sulfur (e.g., 20 ml of medium in 120 ml serum bottles containing 0.6 g sulfur) under a stream of H2-CO2 (4:1, v/v) gas mixture and seal with butyl rubber stoppers. Aseptically and anaerobically add the sterile Na2S x 9H2O solution. Pressurize the inoculated bottle to 200 kPa H2-CO2 (4:1, v/v).


## Trace mineral solution

1.5	g	Nitrilotriacetic acid	
0.5	g	MnSO4 x n H2O	
0.5	g	CoSO4 x 7 H2O	
0.18	g	ZnSO4 x 7 H2O	
0.01	g	CuSO4 x 5 H2O	
0.02	g	AlK(SO4)2	
0.01	g	H3BO3	
0.01	g	Na2MoO4 x 2 H2O	
0.01	g	SrCl2 x 6 H2O	
0.01	g	NaBr	
0.01	g	KI	
1000	ml	Distilled water	

1. Dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH. 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