# 1098: ARCHAEOGLOBUS INFECTUS MEDIUM


## Main sol. 1098

35	g	Sea Salt (SIGMA)	
0.1	g	NH4Cl	
0.05	g	KH2PO4	
1.6	g	Na-acetate	
0.1	g	Yeast extract	
1	ml	**Wolfe's mineral elixir**	
0.5	ml	Sodium resazurin (0.1% w/v)	
2	g	NaHCO3	
0.14	g	2-Mercaptoethanesulfonic acid (coenzyme M)	
1	ml	**Seven vitamins solution**	
2.5	g	Na2S2O3 x 5 H2O	
0.3	g	DL-Dithiothreitol	
1000	ml	Distilled water	

1. Dissolve ingredients (except bicarbonate, coenzyme M, vitamins, thiosulfate, and dithiothreitol), then sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under the same gas atmosphere into anoxic serum vials (e.g., Balch-type tubes) to 30% of their volume and autoclave. Add coenzyme M, vitamins, thiosulfate, and dithiothreitol 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 atmosphere. The vitamin and dithiothreitol solutions should be sterilized by filtration. Adjust the pH of the complete medium to 6.5.
2. After inoculation pressurize culture vessels to 1.5 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture.


## Wolfe's mineral elixir

30	g	MgSO4 x 7 H2O	
5	g	MnSO4 x H2O	
10	g	NaCl	
1	g	FeSO4 x 7 H2O	
1.8	g	CoCl2 x 6 H2O	
1	g	CaCl2 x 2 H2O	
1.8	g	ZnSO4 x 7 H2O	
0.1	g	CuSO4 x 5 H2O	
0.18	g	AlK(SO4)2 x 12 H2O	
0.1	g	H3BO3	
0.1	g	Na2MoO4 x 2 H2O	
2.8	g	(NH4)2Ni(SO4)2 x 6 H2O	
0.1	g	Na2WO4 x 2 H2O	
0.1	g	Na2SeO4	
1000	ml	Distilled water	

1. First adjust pH to 1.0 with diluted H2SO4, then add and dissolve the salts.


## Seven vitamins solution

100	mg	Vitamin B12	
80	mg	p-Aminobenzoic acid	
20	mg	D-(+)-biotin	
200	mg	Nicotinic acid	
100	mg	Calcium pantothenate	
300	mg	Pyridoxine hydrochloride	
200	mg	Thiamine-HCl x 2 H2O	
1000	ml	Distilled water