# 926: ALKALIPHILIC THERMOCOCCUS MEDIUM


## Main sol. 926

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	
0.32	g	NaHCO3	
1	g	K2HPO4	
0.05	mg	CaCl2 x 2 H2O	
15	mg	KI	
0.03	g	H3BO3	
20	ml	**Modified Wolin's mineral solution II**	
2000	ml	Distilled water	

1. Prepare the medium anaerobically under nitrogen. Do not adjust the pH.
2. Prepare separate anaerobic stock solutions of, Casamino acids(10%), Yeast extract (10%), and glycine (2M = 150 g/l). A 0.5M polysulphide solution is prepared by dissolving 12.0 g Na2S x 9 H2O in oxygen free water, followed by adding 1.6 g sulphur - the solution will be dark yellow.


## Modified Wolin's mineral solution II

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	
0.025	g	NiCl2 x 6 H2O	
0.3	mg	Na2SeO3 x 5 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.