# 951: METHYLOPHAGA SULFIDOVORANS MEDIUM


## Main sol. 951

15	g	NaCl	
0.5	g	(NH4)2SO4	
0.33	g	CaCl2 x 6 H2O	
0.2	g	KCl	
1	g	MgSO4 x 7 H2O	
0.02	g	KH2PO4	
2	g	Na2CO3	
1	mg	FeSO4 x 7 H2O	
1	ml	**Trace element solution SL10**	
1	ml	**Vitamin solution**	
10	ml	Methanol	
15	g	Agar	
998	ml	Distilled water	

1. Prepare the medium without methanol and the vitamin solution. Adjust pH to 7.5 (±0.3) with 2 N HCl. Liquid media is prepared in either Hungate tubes (5ml medium) or 100ml serum bottles (10ml medium) sealed with rubber stoppers.
2. Sterilize by autoclaving. After autoclaving add the methanol (filter sterilised) and the vitamin solution (filter sterilised) from sterile stock solutions.


## Trace element solution SL10

5	ml	HCl (concentrated)	
1.5	g	FeCl2 x 4 H2O	
0.07	g	ZnCl2	
0.2	g	MnCl2 x 4 H2O	
6	mg	H3BO3	
0.19	g	CoCl2 x 6 H2O	
2	mg	CuCl2 x 2 H2O	
0.024	g	NiCl2 x 6 H2O	
0.036	g	Na2MoO4 x 2 H2O	
1000	ml	Distilled water	


## Vitamin solution

20	mg	Biotin	
200	mg	Nicotinic acid	
100	mg	Thiamine	
100	mg	p-Aminobenzoic acid	
50	mg	Pantothenate	
500	mg	Pyridoxine hydrochloride	
10	mg	Riboflavine	
10	mg	Vitamin B12	
1000	ml	Distilled water