# 347a: SYNTROPHOMONAS ZEHNDERI MEDIUM


## Main sol. 347a

921	ml	**Solution A**	
30	ml	**Solution B**	
1	ml	**Solution C**	
25	ml	**Solution D**	
10	ml	**Solution E**	
10	ml	**Solution F**	
10	ml	**Solution G**	

1. Dissolve ingredients of solution A, adjust pH to 7.0, then sparge with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Distribute under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Solution B is autoclaved separately under 80% N2 and 20% CO2 gas atmosphere. Solution C is prepared under 100% N2 gas and sterilized by filtration. Solutions D, E, F and G are autoclaved under 100% N2 gas. To complete the medium, appropriate amounts of the solutions B to G are added to solution A in the sequence indicated. Adjust pH of complete medium to 7.2, if necessary.
2. Note: Solutions D and F lead to a white precipitate which makes the medium very cloudy. Alternatively, solutions D and F can be replaced with 10 ml/l of a 14% (w/v) solution of Na-Caproate, which does not precipitate.


## Solution A

0.5	g	KH2PO4	
0.33	g	MgCl2 x 6 H2O	
0.4	g	NaCl	
0.4	g	NH4Cl	
0.05	g	CaCl2 x 2 H2O	
1	ml	**Trace element solution SL-10**	
2.8	g	Na2SO4	
15	g	PIPES (SIGMA)	
0.5	ml	Sodium resazurin (0.1% w/v)	
920	ml	Distilled water	


## Solution B

1.5	g	Na2CO3	
30	ml	Distilled water	


## Solution C

1	ml	**Seven vitamins solution**	


## Solution D

2.78	g	Na-laurate	
25	ml	Distilled water	


## Solution E

1.84	g	CaCl2 x 2 H2O	
10	ml	Distilled water	


## Solution F

0.3	g	L-Cysteine HCl x H2O	
10	ml	Distilled water	


## Solution G

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.


## Seven vitamins solution

100	mg	Vitamin B12	
80	mg	p-Aminobenzoic acid	
20	mg	D-(+)-biotin	
200	mg	Nicotinic acid	
100	mg	Calcium pantothenate	
300	mg	Pyridoxine hydrochloride	
200	mg	Thiamine-HCl x 2 H2O	
1000	ml	Distilled water