# J335: JCM MEDIUM No. 335


## Main sol. J335

0.78	g	K2HPO4	
0.75	g	KH2PO4	
0.04	g	Na3-EDTA	
0.01	g	FeSO4 x H2O	
0.25	g	MgSO4 x H2O	
0.03	g	CaCl2 x H2O	
0.2	g	NaCl	
27	mg	NH4Cl	
2.8	g	Na2SO4	
0.16	g	Sodium acetate	
10	ml	**Trace vitamins**	
10	ml	**Trace element solution**	
0.25	g	L-Cysteine HCl x H2O	
1000	ml	Distilled water	

1. Adjust pH to 5.5. Fill the gas phase with H2-CO2 (80:20).


## 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	


## 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.