# 374: METHANOSPIRILLUM SK MEDIUM


## Main sol. 374

0.5	g	KH2PO4	
0.4	g	MgSO4 x 7 H2O	
0.4	g	NaCl	
0.25	g	NH4Cl	
0.05	g	CaCl2 x 2 H2O	
1	ml	**Trace element solution SL-10**	
1	ml	**Selenite-tungstate solution**	
1	g	Na-acetate	
0.5	ml	Sodium resazurin (0.1% w/v)	
1	g	Na2CO3	
30	ml	Isopropyl alcohol (25% v/v)	
0.5	g	Na2S x 9 H2O	
970	ml	Distilled water	

1. Dissolve ingredients except carbonate, isopropyl alcohol and sulfide. Sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add and dissolve bicarbonate, then dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add isopropyl alcohol and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Prior to use check pH of complete medium and adjust to 7.0 - 7.2, if necessary.


## 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