# J900: M13a MEDIUM


## Main sol. J900

0.25	g	Peptone	
0.25	g	Yeast extract	
0.25	g	Glucose	
10	ml	**Vitamin solution**	
20	ml	**Modified Hutner's basal salts**	
50	ml	Tris-HCl buffer (pH 7.5, 0.1 M)	
500	ml	Sea water (artificial)	
420	ml	Distilled water	

1. For agar plates, increase pH of Tris-HCl buffer to 8.5.


## Vitamin solution

0.2	mg	Vitamin B12	
4	mg	Biotin	
10	mg	Thiamine-HCl x 2 H2O	
10	mg	Calcium pantothenate	
1	mg	Folic acid	
10	mg	Riboflavin	
10	mg	Nicotinamide	
10	mg	p-Aminobenzoic acid	
10	mg	Pyridoxine hydrochloride	
1000	ml	Distilled water	

1. Filter-sterilize.


## Modified Hutner's basal salts

10	g	Nitrilotriacetic acid (NTA)	
29.7	g	MgSO4 x 7 H2O	
3.335	g	CaCl2 x 2 H2O	
9.25	mg	(NH4)6Mo7O24 x 4 H2O	
99	mg	FeSO4 x 7 H2O	
50	ml	**Metals ''44''**	
950	ml	Distilled water	

1. Dissolve nitrilotriacetic acid and adjust pH to 7.0 with KOH (about 7.3 g). Then proceed to add salts. Readjust pH to 6.8 with NaOH or H2SO4.


## Metals ''44''

250	mg	Na2-EDTA	
1095	mg	ZnSO4 x 7 H2O	
500	mg	FeSO4 x 7 H2O	
154	mg	MnSO4 x n H2O	
39.2	mg	CuSO4 x 5 H2O	
24.8	mg	Co(NO3)2 x 6 H2O	
17.7	mg	Na2B4O7 x 10 H2O	
1000	ml	Distilled water