# 1249: GEOALKALIBACTER SUBTERRANEUS MEDIUM


## Main sol. 1249

17	g	NaCl	
2.55	g	NaNO3	
4.5	g	MgCl2 x 6 H2O	
0.35	g	CaCl2 x 2 H2O	
1	g	NH4Cl	
0.08	g	KH2PO4	
1	ml	**Trace element solution SL-10**	
1	ml	**Selenite-tungstate solution**	
1.5	g	Na2CO3	
1	g	Na-acetate	
3	g	Yeast extract	
1000	ml	Distilled water	

1. Dissolve ingredients (except carbonate, acetate and yeast extract), then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Prior to inoculation add acetate and yeast extract from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. The pH of the complete medium should be at 7.0 - 7.2.


## Trace element solution SL-10

10	ml	HCl (25%)	
1.5	g	FeCl2 x 4 H2O	
70	mg	ZnCl2	
100	mg	MnCl2 x 4 H2O	
6	mg	H3BO3	
190	mg	CoCl2 x 6 H2O	
2	mg	CuCl2 x 2 H2O	
24	mg	NiCl2 x 6 H2O	
36	mg	Na2MoO4 x 2 H2O	
990	ml	Distilled water	

1. First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml.


## Selenite-tungstate solution

0.5	g	NaOH	
3	mg	Na2SeO3 x 5 H2O	
4	mg	Na2WO4 x 2 H2O	
1000	ml	Distilled water