# J1104: HYPNOCYCLICUS MEDIUM


## Main sol. J1104

0.02	g	Na2SO4	
1	g	KH2PO4	
0.1	g	NH4Cl	
20	g	NaCl	
3	g	MgCl2 x 6 H2O	
0.5	g	KCl	
0.15	g	CaCl2 x 2 H2O	
1	ml	**FeCl2 solution**	
1	ml	**Trace element solution**	
1	mg	Resazurin	
950	ml	Distilled water	

1. Mix components thoroughly and autoclave under a N2-CO2 (80:20, v/v) atmosphere. Aseptically and anaerobically add the following solutions (autoclaved or *filter-sterilized): 

30	ml	NaHCO3 (*, 8%)	
1	ml	**Vitamin solution** (*)	
5	ml	Yeast extract (10%)	
5	ml	Maltose (1 M)	
10	ml	Na2S x 9 H2O (5%)	

2. Readjust pH to 6.8 - 7.2, if necessary.


## FeCl2 solution

10	ml	HCl (7.7 M, 25%)	
1.5	g	FeCl2 x 4 H2O	
990	ml	Distilled water	


## Trace element solution

70	mg	ZnCl2	
100	mg	MnCl2 x 4 H2O	
6	mg	H3BO3	
190	mg	CoCl2 x 6 H2O	
2	mg	CuCl2 x 2 H2O	
24	mg	NiCl2 x 6 H2O	
36	mg	Na2MoO4 x 2 H2O	
1000	ml	Distilled water	


## Vitamin solution

50	mg	Vitamin B12	
50	mg	p-Aminobenzoic acid	
10	mg	Biotin	
100	mg	Nicotinic acid	
25	mg	DL-Calcium pantothenate	
250	mg	Pyridoxine hydrochloride	
50	mg	Thiamine HCl	
1000	ml	Distilled water