# 1571: MICROBACTER MEDIUM


## Main sol. 1571

0.41	g	KH2PO4	
0.53	g	Na2HPO4 x 2 H2O	
0.3	g	NH4Cl	
0.3	g	NaCl	
0.1	g	MgCl2 x 6 H2O	
0.01	g	CaCl2 x 2 H2O	
2	g	Yeast extract	
0.36	g	Na2S x 9 H2O	
0.5	g	Resazurin	
1	ml	**Trace element solution SL-10**	
1000	ml	Distilled water	
0.5	g	L-Cysteine	

1. Dissolve in 1 liter, boil under N2, add 0.5 g of L-cysteine and adjust pH to 6.4 (should be pH 5.5 after autoclaving). Disperse in tubes and bottles (leave 50% of container volume as gas phase). Change to gas mixture N2/CO2; 80/20%) and autoclave for 20 minutes at 121°C.
2. To 100 ml sterile medium add aseptically 

0.5	ml	**Vitamin solution**	
0.5	ml	**Selenite-tungstate solution**	


## Trace element solution SL-10

10	ml	HCl (25%)	
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. First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml.


## Vitamin solution

2	mg	Biotin	
2	mg	Folic acid	
10	mg	Pyridoxine hydrochloride	
5	mg	Thiamine-HCl x 2 H2O	
5	mg	Riboflavin	
5	mg	Nicotinic acid	
5	mg	D-Calcium pantothenate	
0.1	mg	Vitamin B12	
5	mg	p-Aminobenzoic acid	
5	mg	Lipoic acid	
1000	ml	Distilled water	


## Selenite-tungstate solution

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