# J907: GEOBACTER MEDIUM


## Main sol. J907

0.136	g	KH2PO4	
0.535	g	NH4Cl	
0.204	g	MgCl2 x 6 H2O	
0.147	g	CaCl2 x 2 H2O	
0.82	g	Sodium acetate	
0.142	g	Na2SO4	
10	ml	**Trace element solution**	
10	ml	**Trace vitamins**	
1	ml	**Se/W solution**	
920	ml	Distilled water	

1. Mix components thoroughly and autoclave under a N2-CO2 (80:20, v/v) gas mixture. After cooling, add aseptically and anaerobically the following solutions to the medium: 

30	ml	NaHCO3 (filter-sterilized, 8%)	
50	ml	**100 mM Fe NTA solution** (100 mM)	

2. Adjust pH to 7.0 - 7.2, if necessary.


## 100 mM Fe NTA solution

1.64	g	NaHCO3	
2.75	g	Nitrilotriacetic acid trisodium salt monohydrate	
2.7	g	FeCl3 x 6 H2O	

1. Firstly, dissolve NaHCO3 in 80 ml distilled water. Then, add NTA and FeCl3 x 6H2O to the solution and bring volume to 100 ml with distilled water. Adjust pH to 7.0 with NaOH. Filter-sterilize and store under a N2 atmosphere.
2. Comment:
3. 100 mM Fe NTA solution can be replaced with Iron(III)citrate solution (see Medium No. 1014, autoclaved and stored under N2).


## Trace element solution

12.8	g	Nitrilotriacetic acid	
1.35	g	FeCl3 x 6 H2O	
0.1	g	MnCl2 x 4 H2O	
0.024	g	CoCl2 x 6 H2O	
0.1	g	CaCl2 x 2 H2O	
0.1	g	ZnCl2	
0.025	g	CuCl2 x 2 H2O	
0.01	g	H3BO3	
1	g	NaCl	
0.12	g	NiCl2 x 6 H2O	
0.026	g	Na2SeO3 x 5 H2O	
1000	ml	Distilled water	

1. Dissolve nitrilotriacetic acid first in 200 ml of distilled water, and adjust pH to 6.5 with KOH. Then add the remaining salts and adjust volume to 1.0 L. Readjust pH to 7.0 with KOH.
2. Comment: For strain JCM 10134, incubate with a 10-20% inoculum.


## 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	


## Se/W solution

4	mg	Na2SeO3 x 5 H2O	
4	mg	Na2WO4 x 2 H2O	
1000	ml	Distilled water