# 1801: SULFURIMONAS MARISNIGRI MEDIUM


## Main sol. 1801

1001	ml	**Solution A**	
15	ml	**Solution B**	
2	ml	**Solution C**	
2	ml	**Solution D**	
2	ml	**Solution E**	
3	ml	**Solution F**	
1	ml	**Solution G**	

1. Dissolve ingredients of solution A (except MnO2), then sparge with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Distribute solution A under the same gas atmosphere into anoxic Hungate-type tubes or serum vials containing the appropriate amount of MnO2 and autoclave at 121°C for 15 min. Solution B is autoclaved separately under an 80% N2 and 20% CO2 gas atmosphere, and solutions C, D, and E  are autoclaved under a 100% N2 gas atmosphere. Solutions F and G are prepared under 100% N2 gas atmosphere and sterilized by filtration. To complete the medium appropriate amounts of solutions B to G are added to the sterile solution A in the sequence as indicated. The final pH of the complete medium should be 7.5.


## Solution A

17.7	g	NaCl	
3.2	g	MgCl2 x 6 H2O	
0.4	g	CaCl2 x 2 H2O	
0.7	g	KCl	
2.7	g	MgSO4 x H2O	
1	ml	**Trace element solution SL-10**	
0.5	g	MnO2	
1000	ml	Distilled water	


## Solution B

0.75	g	Na2CO3	
15	ml	Distilled water	


## Solution C

0.01	g	NH4Cl	
2	ml	Distilled water	


## Solution D

0.01	g	K2HPO4	
2	ml	Distilled water	


## Solution E

0.16	g	Na-acetate	
2	ml	Distilled water	


## Solution F

0.75	g	Na2S2O3 x 5 H2O	
3	ml	Distilled water	


## Solution G

1	ml	**Wolin's vitamin solution (10x)**	


## 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.


## Wolin's vitamin solution (10x)

20	mg	Biotin	
20	mg	Folic acid	
100	mg	Pyridoxine hydrochloride	
50	mg	Thiamine HCl	
50	mg	Riboflavin	
50	mg	Nicotinic acid	
50	mg	Calcium D-(+)-pantothenate	
1	mg	Vitamin B12	
50	mg	p-Aminobenzoic acid	
50	mg	(DL)-alpha-Lipoic acid	
1000	ml	Distilled water