# 1291: CHTHONOMONAS MEDIUM


## Main sol. 1291

0.5	g	Gellan Gum (Phytagel)	
4	g	(NH4)2SO4	
0.45	g	K2HPO4	
0.15	ml	CaCl2 x 2 H2O	
1	ml	**Trace element solution SL-11**	
1	g	MgCl2 x 6 H2O	
0.01	g	Yeast extract	
1	ml	**Seven vitamins solution**	
1000	ml	Distilled water	

1. Add gellan gum and boil to dissolve. Then add the remaining ingredients (except magnesium chloride, yeast extract and vitamins), adjust the pH value to 5.0 to 5.3 and fill the medium under an air atmosphere into tubes of the Hungate type or similar vials up to 30% their volume. After autoclaving add magnesium chloride, yeast extract and vitamins from steril stock solutions. Vitamins should be sterilized by filtration.
2. Note: Incubation with shaking may improve cell yield.


## 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. 


## 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