# 498: PSEUDOTHERMOTOGA MEDIUM


## Main sol. 498

0.5	g	Yeast extract (BD Bacto)	
15	ml	**Modified Wolin's mineral solution**	
3.46	g	NaCl	
0.77	g	Na2-EDTA	
0.88	g	MgSO4 x 7 H2O	
0.69	g	MgCl2 x 6 H2O	
0.14	g	CaCl2 x 2 H2O	
0.08	g	KCl	
12.5	mg	NaBr	
4	ml	H3BO3 (0.1% w/v)	
3	ml	(NH4)2Ni(SO4)2 (0.1% w/v)	
1.9	ml	SrCl2 x 6 H2O (0.1% w/v)	
0.1	ml	KI (0.01% w/v)	
0.5	ml	Sodium resazurin (0.1% w/v)	
0.5	g	KH2PO4	
5	g	Starch (soluble)	
0.5	g	Na2S x 9 H2O	
1000	ml	Distilled water	

1. Dissolve ingredients (except hydrogenphosphate, starch and sulfide) and adjust pH to 7.0. Sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic, dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% volume and autoclave. Add hydrogenphosphate, starch and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Check pH of the complete medium and adjust to 7.0 - 7.2, 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.