# J920: HYDROGENISPORA MEDIUM


## Main sol. J920

0.2	g	Yeast extract (BD-Difco)	
0.14	g	KH2PO4	
0.2	g	MgCl2 x 6 H2O	
0.15	g	CaCl2 x 2 H2O	
0.54	g	NH4Cl	
1	ml	**Trace vitamins solution**	
1	ml	**Trace element solution**	
1	ml	**Se/W solution**	
2.5	g	NaHCO3	
1	mg	Resazurin	
1000	ml	Distilled water	

1. Mix components, except NaHCO3, and adjust pH to 6.5. Bring to a boil for 5-10 sec, cool down under a stream of a N2-CO2 (80:20 v/v) gas mixture and add NaHCO3 to the solution. Distribute the medium into culture vessels (e.g., 20 ml medium in 60 ml serum bottle) under the same gas mixture and seal with butyl rubber stoppers. After autoclaving, stand the medium overnight. Then, add per liter the following solutions: 

10	ml	Glucose (1.0 M)	
6	ml	L-Cysteine HCl x H2O (5%)	
6	ml	Na2S x 9 H2O (5%)	


## Trace vitamins solution

4.9	mg	Biotin	
8.8	mg	Folic acid	
4.1	mg	Pyridoxine hydrochloride	
6.7	mg	Thiamine HCl	
7.5	mg	Riboflavin	
2.5	mg	Nicotinic acid	
9.5	mg	DL-Calcium pantothenate	
27.1	mg	Vitamin B12	
2.7	mg	p-Aminobenzoic acid	
4.1	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	
0.024	g	Na2MoO4 x 2 H2O	
0.12	g	NiCl2 x 6 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 bring volume to 1.0 L. Readjust pH to 7.0 with KOH.


## Se/W solution

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