# 1367: TELMATOCOLA MEDIUM


## Main sol. 1367

0.1	g	KH2PO4	
0.1	g	(NH4)2SO4	
0.1	g	MgSO4 x 7 H2O	
0.02	g	CaCl2 x 2 H2O	
1	ml	**Hutner's salts**	
1	ml	**Vitamin solution**	
0.1	g	Yeast extract	
0.1	g	Glucose	
1000	ml	Distilled water	

1. Prepare the liquid medium without Staley´s vitamin solution and glucose. Adjust to pH 4.8 - 5.5. Dispense the medium into serum bottles. The gas phase is air:CO2 (95:5) and the liquid:gas phase should be 1:9. Sterilize by autoclaving. After autoclaving add the Staley´s vitamin solution (filter sterilised) and glucose (autoclaved) from sterile stock solutions.


## 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.


## "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.


## Vitamin solution

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. Filter-sterilized.