# J764: HEPES PEPTONE-YEAST EXTRACT MEDIUM


## Main sol. J764

2.38	g	HEPES	
30	g	NaCl	
2.7	g	MgSO4 x 7 H2O	
4.3	g	MgCl2 x 6 H2O	
0.25	g	NH4Cl	
0.5	g	KCl	
0.14	g	CaCl2 x 2 H2O	
0.14	g	K2HPO4	
2	g	Bacto peptone (BD-Difco)	
0.2	g	Yeast extract	
1	ml	**Trace elements**	
1	ml	Fe(NH4)2(SO4)2 x 6 H2O (in 0.037% HCl, 0.2%)	
1000	ml	Distilled water	

1. Adjust pH to 7.25. Autoclave and after cooling, add the following solution (filter-sterilized): 

10	ml	**Trace vitamins**	


## 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