# J1302: SEA SALTS YTG MEDIUM


## Main sol. J1302

1. Mix components and adjust pH to 7.0. Dispense the medium into culture vessels under a N2 gas atmosphere (e.g., 10 ml medium in Hungate tubes), seal with butyl rubber stoppers and autoclave. After cooling, aseptically and anaerobicallly add per liter the following solutions (filter-sterilized): 

0.5	g	Yeast extract	
1	g	Tryptone	
2.5	g	Glucose	
30	g	Sea Salt (Sigma-Aldrich)	
1	ml	**Trace minerals**	
1	ml	Na2SeO3 (0.1%)	
0.2	g	L-Cysteine HCl x H2O	
0.5	mg	Resazurin	
1000	ml	Distilled water	
10	ml	**Trace vitamins**	
5	ml	Sodium Thiosulfate (1M)	


## 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