# J621: HYL MEDIUM


## Main sol. J621

4.76	g	HEPES	
20	g	NaCl	
4	g	Na2SO4	
4.3	g	MgCl2 x 6 H2O	
0.25	g	NH4Cl	
0.34	g	KCl	
0.14	g	CaCl2 x 2 H2O	
0.14	g	K2HPO4	
1	g	Yeast extract	
4.5	ml	Sodium lactate (50%)	
1	mg	Resazurin	
1000	ml	Distilled water	

1. Autoclave and cool down to room temperature while flushing with N2. Then add the following solutions from sterile anaerobic stocks: 

1	ml	**Trace elements**	
1	ml	Fe(NH4)2(SO4)2 x 6 H2O (in 0.037% HCl, 0.2%)	
10	ml	**Trace vitamins**	
4	ml	Na2S x 9 H2O (0.5 M)	

2. Adjust pH to 7.0.


## Trace elements

29	g	Na2-EDTA	
3.26	g	MnSO4 x n H2O	
1.8	g	CoCl2 x 6 H2O	
1	g	ZnSO4 x 7 H2O	
0.11	g	NiSO4 x 6 H2O	
0.1	g	CuSO4 x 5 H2O	
0.1	g	H3BO3	
0.1	g	AlK(SO4)2 x 12 H2O	
0.1	g	Na2MoO4 x 2 H2O	
0.1	g	Na2WO4 x 2 H2O	
0.05	g	Na2SeO3	
1000	ml	Distilled water	

1. First dissolve EDTA x 2Na and adjust pH to 8.0 with KOH. Then add the minerals.


## 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