# 343a: THERMOSIPHO MELANESIENSIS MEDIUM


## Main sol. 343a

5	g	Starch (soluble)	
0.5	g	KH2PO4	
15	ml	**Modified Wolin's mineral solution**	
2	ml	NiCl2 x 6 H2O (0.1% w/v)	
20	g	NaCl	
250	ml	**Artificial sea water**	
2	g	Yeast extract (OXOID)	
0.5	ml	Sodium resazurin (0.1% w/v)	
0.5	g	L-Cysteine HCl x H2O	
0.5	g	Na2S x 9 H2O	
750	ml	Distilled water	

1. Dissolve ingredients (except sulfide and cysteine) and adjust pH to 6.5. Boil medium for 1 min, then cool to room temperature under 100% N2 gas atmosphere. Dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% of volume and autoclave. Add sulfide and cysteine from sterile anoxic stock solutions prepared under 100% N2 gas. Adjust pH of complete medium to 6.5, if necessary.


## Modified Wolin's mineral solution

1.5	g	Nitrilotriacetic acid	
3	g	MgSO4 x 7 H2O	
0.5	g	MnSO4 x H2O	
1	g	NaCl	
0.1	g	FeSO4 x 7 H2O	
0.18	g	CoSO4 x 7 H2O	
0.1	g	CaCl2 x 2 H2O	
0.18	g	ZnSO4 x 7 H2O	
0.01	g	CuSO4 x 5 H2O	
0.02	g	AlK(SO4)2 x 12 H2O	
0.01	g	H3BO3	
0.01	g	Na2MoO4 x 2 H2O	
0.03	g	NiCl2 x 6 H2O	
0.3	mg	Na2SeO3 x 5 H2O	
0.4	mg	Na2WO4 x 2 H2O	
1000	ml	Distilled water	

1. First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Adjust final to pH 7.0 with KOH.


## Artificial sea water

27.7	g	NaCl	
7	g	MgSO4 x 7 H2O	
5.5	g	MgCl2 x 6 H2O	
0.65	g	KCl	
0.1	g	NaBr	
30	mg	H3BO3	
15	mg	SrCl2 x 6 H2O	
10	mg	Citric acid	
0.05	mg	KI	
2.25	g	CaCl2 x 2 H2O	
1000	ml	Distilled water