# J754: MJ MEDIUM FOR DEFFERIBACTER SP. STRAIN NEJINEJI


## Main sol. J754

30	g	NaCl	
6.5	g	MgCl2 x 6 H2O	
0.5	g	CaCl2 x 2 H2O	
0.33	g	KCl	
0.25	g	NH4Cl	
0.14	g	K2HPO4	
4	mg	MnCl2 x 4 H2O	
4	mg	Na2SeO3 x 5 H2O	
2	mg	NiCl2 x 6 H2O	
2	mg	CoCl2 x 6 H2O	
0.5	mg	ZnCl2	
0.4	mg	Na2MoO4 x 2 H2O	
0.25	mg	FeCl2 x 4 H2O	
0.1	mg	Na2WO2 x 2 H2O	
0.1	mg	H3BO3	
10	g	Sulfur	
0.5	mg	Resazurin	
1000	ml	Distilled water	

1. Mix components except sulfur and autoclave under a N2-CO2 (4:1, v/v) gas mixture. After cooling, add the following solutions from anaerobic stocks (filter-sterilized): 

1	ml	**Trace vitamins**	
12.5	ml	NaHCO3 (8%)	

2. Separately steam sulfur for 3 hr on each of 3 successive days.
3. To complete the medium, distribute the solution into sterile culture vessels containing appropriate amount of sulfur (e.g., 5 ml solution and 0.05 g sulfur in Hungate tubes) under a H2-CO2 (4:1, v/v) gas mixture and seal with butyl rubber stoppers. Then, add Na2S x 9H2O solution from an anaerobic stock (final 0.1 g/L). After inoculation, pressurize the inoculated culture vessels to 200 kPa H2-CO2 (4:1, v/v).


## Trace vitamins

2	mg	Biotin	
2	mg	Folic acid	
10	mg	Pyridoxine hydrochloride	
5	mg	Thiamine HCl	
5	mg	Riboflavin	
5	mg	Nicotinic acid	
5	mg	Calcium pantothenate	
0.1	mg	Vitamin B12	
5	mg	p-Aminobenzoic acid	
5	mg	Lipoic acid	
1000	ml	Distilled water