# 113: THIOBACILLUS DENITRIFICANS MEDIUM


## Main sol. 113

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

1. Solution A is sterilized by autoclaving at 121°C for 15 min under a 100% N2 gas atmosphere. Solutions B and D are sterilized by filtration under a 100% N2 gas atmosphere. Solution C is sterilized by autoclaving under an atmosphere of 80% N2 and 20% CO2 gas mixture. Appropriate amounts of solutions B to D are added to the sterile solution A in the sequence as indicated. Check the pH of the complete medium and adjust it to 7.0 - 7.2, if necessary.


## Solution A

2	g	KH2PO4	
2	g	KNO3	
1	g	NH4Cl	
0.8	g	MgSO4 x 7 H2O	
2	ml	**Trace element solution SL-4**	
960	ml	Distilled water	


## Solution B

5	g	Na2S2O3 x 5 H2O	
20	ml	Distilled water	


## Solution C

1	g	Na2CO3	
20	ml	Distilled water	


## Solution D

20	mg	FeSO4 x 7 H2O	
10	ml	H2SO4 (0.1 N)	


## Trace element solution SL-4

0.5	g	Na2-EDTA	
0.2	g	FeSO4 x 7 H2O	
0.1	g	ZnSO4 x 7 H2O	
0.03	g	MnCl2 x 4 H2O	
0.3	g	H3BO3	
0.2	g	CoCl2 x 6 H2O	
0.01	g	CuCl2 x 2 H2O	
0.02	g	NiCl2 x 6 H2O	
0.03	g	Na2MoO4 x 2 H2O	
1000	ml	Distilled water	

1. First dissolve EDTA in distilled water and adjust pH to 7.0 using 2 N NaOH; then add other compounds.