# 770: SULFOPHOBOCOCCUS ZILLIGII MEDIUM


## Main sol. 770

970	ml	**Solution A**	
10	ml	**Solution B**	
10	ml	**Solution C**	
5	ml	**Solution D**	
10	ml	**Solution E**	

1. Dissolve ingredients of solution A except carbonate, sparge with 100% N2 gas for 30 - 45 min to make it anoxic, then add carbonate and adjust pH to 9.0 - 9.3. Dispense under 100% N2 gas atmosphere into anoxic Hungate-type tubes or serum vials (e.g., 30 ml medium per 100 ml bottle) and autoclave. Solutions B and C are autoclaved separately under 100% N2 gas atmosphere. Solutions D and E are prepared under 100% N2 gas and sterilized by filtration. To complete the medium add appropriate amounts of solutions B to E to the sterile solution A in the sequence as indicated. Adjust pH of complete medium to 7.5.
2. Note: Addition of 10 - 20 mg sodium dithionite per liter (e.g. from 5% (w/v) solution, freshly prepared under N2 and filter-sterilized) may stimulate growth at the beginning.
3. For transfers use 10% (v/v) inoculum.


## Solution A

32	mg	Na2-EDTA	
66	mg	CaCl2 x 2 H2O	
31	mg	MgSO4 x 7 H2O	
31	mg	KCl	
2.1	ml	ZnCl2 (0.1% w/v)	
2.3	ml	MnSO4 x H2O (0.1% w/v)	
1.8	ml	Na2B4O7 x 10 H2O (0.1% w/v)	
1.5	g	Glycine	
0.5	ml	Sodium resazurin (0.1% w/v)	
230	mg	Na2CO3	
965	ml	Distilled water	


## Solution B

1	g	Yeast extract	
10	ml	Distilled water	


## Solution C

1	g	Trypticase peptone (BD BBL)	
10	ml	Distilled water	


## Solution D

50	mg	FeSO4 x 7 H2O	
5	ml	H2SO4 (0.1 N)	


## Solution E

150	mg	Dithiothreitol (DTT)	
10	ml	Distilled water