# J819: BACILLUS MACYAE MEDIUM


## Main sol. J819

1.2	g	NaCl	
0.3	g	KCl	
0.3	g	NH4Cl	
0.2	g	K2HPO4	
0.3	g	Na2SO4	
0.4	g	MgCl2 x 6 H2O	
0.15	g	CaCl2 x 2 H2O	
0.6	g	NaHCO3	
1	ml	**Trace element solution SL-10**	
0.5	mg	Resazurin	
10	ml	**Trace vitamins**	
1	ml	**Vitamin solution**	
1.1	g	Sodium lactate	
0.8	g	Yeast extract	
0.5	g	KNO3	
1000	ml	Distilled water	

1. Prepare the medium anaerobically under 100% N2. Add vitamins (filter-sterilized), sodium lactate and nitrate from sterile, anaerobic stock solutions. Adjust final pH to 7.4 - 7.8.


## Vitamin solution

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


## Trace element solution SL-10

10	ml	HCl (25%, 7.7 M)	
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	
990	ml	Distilled water	

1. Dissolve FeCl2 x 4H2O first in HCl. Then dilute in distilled water and add the remaining salts.


## 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