# 333: THERMITHIOBACILLUS MEDIUM


## Main sol. 333

0.4	g	NH4Cl	
0.8	g	MgSO4 x 7 H2O	
10	ml	**Trace element solution**	
4	g	KH2PO4	
4	g	K2HPO4	
3	g	K2S4O6	
2	ml	Bromocresol purple (saturated aq. solution)	
10	g	Agar (BD Bacto), for solid medium	
1000	ml	Distilled water	

1. Dissolve ingredients except phosphates and tetrathionate. The phosphates are sterilized separately in 1/10 of medium volume and mixed with the other salts when cool. Tetrathionate is sterilized by filtration and added to the autoclaved medium. Adjust pH of complete medium to 6.9. For agar medium autoclave the phosphates (in 100 ml), other salts (in 300 ml), and agar (in 600 ml) separately.


## Trace element solution

50	g	Na2-EDTA	
11	g	ZnSO4 x 7 H2O	
7.34	g	CaCl2 x 2 H2O	
2.5	g	MnCl2 x 4 H2O	
0.5	g	CoCl2 x 6 H2O	
0.5	g	(NH4)6Mo7O24 x 4 H2O	
5	g	FeSO4 x 7 H2O	
0.2	g	CuSO4 x 5 H2O	
11	g	NaOH	
1000	ml	Distilled water	

1. Dissolve EDTA in distilled water and adjust pH to 7.0 using 2 N NaOH; then add other compounds. Adjust pH of final solution to 6.0 with NaOH.