# 612: DESULFOSARCINA CETONICA MEDIUM


## Main sol. 612

962	ml	**Solution A**	
20	ml	**Solution B**	
10	ml	**Solution C**	
10	ml	**Solution D**	

1. Solution A is sparged with 80% N2 and 20% CO2 gas mixture to reach a pH below 6 (at least 30 min), then distributed under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclaved. Solution B is autoclaved separately under 80% N2 and 20% CO2 gas atmosphere. Solutions C and D are autoclaved under 100% N2 gas atmosphere. To complete the medium appropriate amounts of solutions B to D are added to the sterile solution A in the sequence as indicated. Final pH of the medium should be 7.2 – 7.4.


## Solution A

0.7	g	KH2PO4	
0.3	g	NH4Cl	
2.8	g	Na2SO4	
1.7	g	MgCl2 x 6 H2O	
0.05	g	CaCl2 x 2 H2O	
10	g	NaCl	
1	ml	**Trace element solution SL-10**	
0.5	ml	Sodium resazurin (0.1% w/v)	
960	ml	Distilled water	


## Solution B

1	g	Na2CO3	
20	ml	Distilled water	


## Solution C

1.2	g	Na-butyrate	
10	ml	Distilled water	


## Solution D

0.3	g	Na2S x 9 H2O	
10	ml	Distilled water	


## 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.