# 1544: . PIRELLULA MEDIUM (M30)


## Main sol. 1544

950	ml	**Solution 1**	
50	ml	**Solution 2**	


## Solution 1

20	ml	**Hutner's salts**	
50	ml	Tris-HCl buffer (0.1M, pH 7.5)	
8	g	Gelrite	
1	g	MgCl2	
700	ml	Sea water (Biomaris )	
190	ml	Distilled water	

1. Adjust pH to 7.5, autoclave at 121°C for 20 min.


## Solution 2

2	g	N-Acetylglucosamine	
0.01	g	Na2HPO4 x 2 H2O	
10	ml	**Vitamin solution No.6**	
40	ml	Distilled water	

1. Adjust pH to 7.5, filter-sterilize.


## Vitamin solution No.6

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


## Hutner's 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	Metall salt sol. 44	
950	ml	Distilled water	

1. Dissolve the nitrilotriacetic acid, adjust the pH to 7.0 with KOH (about 7.3 g). Dissolve other salts separately, combine and adjust the pH to 6.8 with NaOH or H2SO4.