# 482: ANAEROBACTER MEDIUM


## Main sol. 482

0.33	g	KH2PO4	
0.33	g	NH4Cl	
0.33	g	MgCl2 x 6 H2O	
0.33	g	CaCl2 x 2 H2O	
0.33	g	KCl	
1	ml	**Trace element solution SL-10**	
1	g	Yeast extract	
0.5	ml	Sodium resazurin (0.1% w/v)	
1.5	g	NaHCO3	
2	g	Sucrose	
0.3	g	L-Cysteine HCl x H2O	
0.3	g	Na2S x 9 H2O	
1000	ml	Distilled water	

1. Dissolve ingredients (except bicarbonate, sucrose, cysteine and sulfide), adjust pH to 7.0, and sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After sterilization add sucrose, cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Adjust pH of complete medium to pH 7.0, 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.