# 484: THIOMICROSPIRA THYASIRAE MEDIUM


## Main sol. 484

100	ml	**Solution A**	
860	ml	**Solution B**	
10	ml	**Solution C**	
20	ml	**Solution D**	
10	ml	**Solution E**	

1. Autoclave solutions A, B and C separately and mix when cool. After autoclaving, add thiosulfate, trace elements and vitamins from sterile stock solutions to complete the medium. Adjust pH of complete medium to 7.3 - 7.6.


## Solution A

7.9	g	Na2HPO4 x 2 H2O	
1.5	g	KH2PO4	
100	ml	Distilled water	

1. Adjust to pH 7.6.


## Solution B

0.4	g	NH4Cl	
0.1	g	MgSO4 x 7 H2O	
25	g	NaCl	
15	g	Agar, for solid medium	
860	ml	Distilled water	


## Solution C

10	ml	**Trace element solution**	


## Solution D

5	g	Na2S2O3 x 5 H2O	
20	ml	Distilled water	


## Solution E

10	ml	**Vitamin solution**	


## Vitamin solution

10	mg	Thiamine-HCl x 2 H2O	
20	mg	Nicotinic acid	
20	mg	Pyridoxine hydrochloride	
10	mg	p-Aminobenzoic acid	
20	mg	Riboflavin	
20	mg	Calcium pantothenate	
1	mg	Biotin	
1	mg	Vitamin B12	
1000	ml	Distilled water	

1. Adjust pH to 7.0.


## 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.