# 1565: ALKALIPHILUS TRANSVAALENSIS MEDIUM


## Main sol. 1565

100	ml	**MJ(-N) synthetic seawater**	
2	g	Yeast extract (BD Bacto)	
2	g	Tryptone (BD Bacto)	
1	g	Na2-tartrate x 2 H2O	
1	g	Na3-citrate	
0.25	g	NaNO3	
0.25	g	(NH4)2SO4	
0.2	g	Ferric citrate monohydrate	
1	g	Na2SO4	
1.25	g	Na2CO3	
2.5	g	NaHCO3	
2	g	Na2S2O3 x 5 H2O	
900	ml	Distilled water	

1. Dissolve ingredients except carbonate, bicarbonate, and thiosulfate, then sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Add and dissolve carbonate and bicarbonate, adjust pH to 10 with KOH, then dispense medium under 100% N2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add thiosulfate to the autoclaved medium from an anoxic stock solution prepared under 100% N2 gas and sterilized by filtration. Adjust the pH of the complete medium to 9.8 - 10.0.


## MJ(-N) synthetic seawater

30	g	NaCl	
0.14	g	K2HPO4	
0.14	g	CaCl2 x 2 H2O	
3.4	g	MgSO4 x 7 H2O	
4.18	g	MgCl2 x 6 H2O	
0.33	g	KCl	
0.5	mg	NiCl2 x 6 H2O	
0.5	mg	Na2SeO3 x 5 H2O	
0.01	g	Fe(NH4)2(SO4)2 x 6 H2O	
10	ml	**Modified Wolin's mineral solution**	
1000	ml	Distilled water	


## 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.