# 778a: THERMOLITHOBACTER MEDIUM


## Main sol. 778a

8.25	g	Na2-9,10-anthraquinone-2,6-disulfonate (TCI A0308)	
0.33	g	KH2PO4	
0.33	g	NH4Cl	
0.33	g	KCl	
0.33	g	MgCl2 x 6 H2O	
1	ml	**Trace element solution SL-10**	
1	ml	**Selenite-tungstate solution**	
0.2	ml	NiCl2 x 6 H2O (0.1% w/v)	
1	g	Yeast extract	
2.5	g	NaHCO3	
0.33	g	CaCl2 x 2 H2O	
1	ml	**Wolin's vitamin solution (10x)**	
1000	ml	Distilled water	

1. Dissolve ingredients (except bicarbonate, calcium chloride and vitamins), boil medium for some minutes to dissolve the anthraquinone, then cool under 80% H2 and 20% CO2 gas atmosphere to room temperature. Add solid bicarbonate and adjust pH to 7.2, dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add calcium chloride and vitamins from sterile anoxic stock solutions prepared under 100% N2 gas atmosphere. Vitamins are sterilized by filtration. Adjust pH of complete medium to 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	


## Wolin's vitamin solution (10x)

20	mg	Biotin	
20	mg	Folic acid	
100	mg	Pyridoxine hydrochloride	
50	mg	Thiamine HCl	
50	mg	Riboflavin	
50	mg	Nicotinic acid	
50	mg	Calcium D-(+)-pantothenate	
1	mg	Vitamin B12	
50	mg	p-Aminobenzoic acid	
50	mg	(DL)-alpha-Lipoic acid	
1000	ml	Distilled water