# 1444: PIRELLULA MEDIUM (M30a)


## Main sol. 1444

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)	
500	ml	**Artifical sea water**	
18	g	Agar	
380	ml	Distilled water	

1. Adjust pH to 7.5, autoclave at 121°C for 20 min.
2. For agar plates increase the pH of the Tris/HCl buffer to 8.5.


## Solution 2

2	g	N-Acetylglucosamine	
0.1	g	Na2HPO4 x 2 H2O	
10	ml	**Vitamin solution (double conc.)**	
40	ml	Distilled water	

1. Adjust pH to 7.5, filter-sterilize, and add to solution 1.


## 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	**"Metals 44"**	
950	ml	Distilled water	

1. Dissolve the nitrilotriacetic acid.
2. Adjust pH to 7.0 with KOH (~7.3 g).
3. Dissolve other salts separately and combine.
4. Adjust pH to 6.8 with NaOH or H2SO4.


## Artifical sea water

23.47	g	NaCl	
3.92	g	Na2SO4	
10.64	g	MgCl2 x 6 H2O	
1.1	g	CaCl2	
192	mg	NaHCO3	
664	mg	KCl	
96	mg	KBr	
26	mg	H3BO3	
24	mg	SrCl2	
3	mg	NaF	
1000	ml	Distilled water	


## Vitamin solution (double conc.)

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

1. Store in the dark and cold (5°C).


## "Metals 44"

250	mg	Na-EDTA	
1095	mg	ZnSO4 x 7 H2O	
500	mg	FeSO4 x 7 H2O	
154	mg	MnSO4 x 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	

1. Dissolve the EDTA and add a few drops of concentrated H2SO4 to retard precipitation of the heavy metal ions.