# J215: PYROBACULUM AEROPHILUM MEDIUM


## Main sol. J215

125	ml	**Marine medium/Synthetic seawater mix solution**	
10	ml	**Trace minerals**	
2	mg	Fe(NH4)2(SO4)2 x 6 H2O	
0.25	g	NH4Cl	
2	mg	(NH4)2Ni(SO4)2 x 6 H2O	
0.1	mg	Na2SeO4	
0.1	mg	Na2WO4 x 2 H2O	
2.2	g	NaHCO3	
0.07	g	KH2PO4	
0.5	g	Yeast extract (BD-Difco)	
1	g	Na2S2O3 x 5 H2O	
865	ml	Distilled water	

1. Adjust pH to 7.0 with 1 N H2SO4, and sterilize by filtration. Dispense the medium into bottles under a N2-CO2-O2 (80:20:1) atmosphere. Pressurize inoculated bottles to 200 kPa N2-CO2-O2 (80:20:1).


## Marine medium/Synthetic seawater mix solution

47.15	g	NaCl	
18.1	g	MgCl2 x 6 H2O	
7	g	MgSO4 x 7 H2O	
3.13	g	CaCl2 x 2 H2O	
3.24	g	Na2SO4	
0.1	g	Na2CO3	
0.1	g	NaBr	
80	mg	KBr	
72	mg	SrCl2 x 6 H2O	
52	mg	H3BO3	
8.1	mg	Na2HPO4	
2.4	mg	NaF	
0.4	mg	Sodium silicate	
0.05	mg	KI	
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.