# 1208: BACILLUS ALKALIDIAZOTROPHICUS MEDIUM


## Main sol. 1208

8	g	NaHCO3	
22	g	Na2CO3	
6	g	NaCl	
1	g	K2HPO4	
0.2	g	NH4Cl	
0.2	g	MgCl2 x 6 H2O	
0.05	g	Yeast extract	
2.8	g	Na2SO4	
1	ml	**Trace element solution SL-11**	
1	ml	**Selenite-tungstate solution**	
10	ml	**Wolin's vitamin solution**	
2	g	Glucose	
1000	ml	Distilled water	

1. Dissolve ingredients except ammonium chloride, magnesium chloride, yeast extract, trace elements, vitamins, boil medium for 1 min, then cool to room temperature under N2 gas atmosphere. Dispense under same gas atmosphere in culture vessels and autoclave. After sterilization add ammonium chloride, magnesium chloride, yeast extract, trace elements, vitamins, from sterile, anoxic stock solution prepared under N2. The vitamin solution should be sterilized by filtration. Adjust pH of final medium to 10.


## 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. 


## Selenite-tungstate solution

0.5	g	NaOH	
3	mg	Na2SeO3 x 5 H2O	
4	mg	Na2WO4 x 2 H2O	
1000	ml	Distilled water	


## Wolin's vitamin solution

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 D-(+)-pantothenate	
0.1	mg	Vitamin B12	
5	mg	p-Aminobenzoic acid	
5	mg	(DL)-alpha-Lipoic acid	
1000	ml	Distilled water