# 818: CHRYSIOGENES MEDIUM


## Main sol. 818

1.2	g	NaCl	
0.3	g	KCl	
0.3	g	NH4Cl	
0.2	g	KH2PO4	
0.3	g	Na2SO4	
0.4	g	MgCl2 x 6 H2O	
0.15	g	CaCl2 x 2 H2O	
1	ml	**Trace element solution SL-11**	
0.5	ml	Sodium resazurin (0.1% w/v)	
0.6	g	NaHCO3	
0.5	g	KNO3	
1	ml	**Wolin's vitamin solution (10x)**	
1	ml	**Seven vitamins solution**	
0.5	g	Na-acetate	
1000	ml	Distilled water	

1. Dissolve ingredients except bicarbonate, nitrate, vitamins and acetate. Sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic, adjust pH to 7.0 - 7.4, then dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add nitrate, vitamins and acetate from sterile anoxic stock solutions prepared under 100% N2 gas and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas atmosphere. Vitamins should be sterilized by filtration. The pH of the complete medium should be adjusted to 7.4 - 7.8, if necessary.


## Trace element solution SL-11

5.2	g	Na2-EDTA x 2 H2O	
1.5	g	FeCl2 x 4 H2O	
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	

1. Dissolve EDTA in 800 ml distilled water, adjust pH to 7 using 2 N NaOH and add ferrous chloride. After ferrous chloride has dissolved add other compounds. Finally adjust pH to 6.0 and bring volume to 1000 ml. 


## Wolin's vitamin solution (10x)

20	mg	Biotin	
20	mg	Folic acid	
100	mg	Pyridoxine hydrochloride	
50	mg	Thiamine HCl	
50	mg	Riboflavin	
50	mg	Nicotinic acid	
50	mg	Calcium D-(+)-pantothenate	
1	mg	Vitamin B12	
50	mg	p-Aminobenzoic acid	
50	mg	(DL)-alpha-Lipoic acid	
1000	ml	Distilled water	


## Seven vitamins solution

100	mg	Vitamin B12	
80	mg	p-Aminobenzoic acid	
20	mg	D-(+)-biotin	
200	mg	Nicotinic acid	
100	mg	Calcium pantothenate	
300	mg	Pyridoxine hydrochloride	
200	mg	Thiamine-HCl x 2 H2O	
1000	ml	Distilled water