# 1505a: PROTEINIVORAX HYDROGENIFORMANS MEDIUM


## Main sol. 1505a

0.3	g	KH2PO4	
0.2	g	MgCl2 x 6 H2O	
0.12	g	MgSO4 x 7 H2O	
0.03	g	CaCl2 x 2 H2O	
50	g	NaCl	
5	g	Tryptone (BD Bacto)	
1	ml	**Trace element solution SL-10**	
1	ml	**Selenite-tungstate solution**	
5	g	Na2CO3	
18	g	NaHCO3	
1	g	**Seven vitamins solution**	
1000	ml	Distilled water	

1. Dissolve ingredients (except sodium carbonate, sodium hydrogencarbonate and vitamins), then sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Add carbonate and hydrogencarbonate, adjust pH to 8.5 - 9.0, dispense medium under 100% N2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After autoclaving add vitamins from an anoxic stock solution prepared under 100% N2 gas and sterilized by filtration.


## 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.


## 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	


## Selenite-tungstate solution

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