# J1017: PFENNIG THERMOFILUM MEDIUM


## Main sol. J1017

0.33	g	NH4Cl	
0.33	g	KH2PO4	
0.33	g	KCl	
0.33	g	CaCl2 x 2 H2O	
0.33	g	MgCl2 x 6 H2O	
1	ml	**Trace element solution**	
10	ml	**Trace vitamins**	
2	g	Glucose	
0.1	g	Yeast extract (Oxoid)	
0.5	g	Trypticase peptone (BD-BBL)	
10	ml	**Archaeal culture filtrate**	
1	mg	Resazurin	
1000	ml	Distilled water	

1. Mix components thoroughly, adjust pH to 6.0 and autoclave under a N2 gas atmosphere. Before inoculation, add per liter the following solution (autoclaved and stored under a N2 gas atmosphere): 

6	ml	Na2S x 9 H2O (neutralized, 5%)	


## Archaeal culture filtrate

1. Cultivate <I>Desulfurococcus kamchatkensis</I> strain 1221n (=JCM 16383) in the above described medium supplemented with 0.6 g/L (final) yeast extract (without the archaeal culture filtrate). After three days of the cultivation, filtrate the grown culture with 0.22 �m (pore size) filter.
2. Comment: For cultivation of JCM 19810, increase amount of yeast extract to 1.0 g/L. Growth may be affected by quality of yeast extract.


## Trace element solution

784	mg	Fe(NH4)2(SO4)2 x 6 H2O	
238	mg	CoCl2 x 6 H2O	
395	mg	(NH4)2Ni(SO4)2 x 6 H2O	
24	mg	Na2MoO4 x 2 H2O	
33	mg	Na2WO4 x 2 H2O	
144	mg	ZnSO4 x 7 H2O	
2	mg	CuCl2 x 2 H2O	
94	mg	Na2SeO4	
6	mg	H3BO3	
99	mg	MnCl2 x 4 H2O	
1000	ml	Distilled water	


## 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