# 1633: V2 MEDIUM


## Main sol. 1633

0.25	g	Peptone (Bacto)	
0.25	g	Yeast extract (Bacto)	
2.38	g	HEPES	
20	ml	**Mineral salts solution**	
1	ml	**Trace element solution**	
0.025	g	NaHCO3	
0.025	g	CaCO3	
0.015	g	CaCl2 x 2 H2O	
0.049	g	MgSO4 x 7 H2O	
0.01	g	KNO3	
1.4	mg	KH2PO4	
0.53	mg	NH4Cl	
15	g	Agar (Bacto)	
730	ml	Double distilled water	

1. Sterilize 20 min./121°C. After cooling to 60°C add: 

8	ml	N-Acetylglucosamine (10% stock solution, 10 % stock solution)	
10	ml	Glucose (2.5%, sterile-filtered)	
5	ml	**Vitamin solution (double conc.)** (medium 621)	

2. Adjust pH to 7.0 with 40% KOH.


## Mineral salts solution

10	g	Nitrilotriacetic acid (NTA)	
29.7	g	MgSO4 x 7 H2O	
3.34	g	CaCl2 x 2 H2O	
12.67	mg	Na2MoO7O4 x 2 H2O	
99	mg	FeSO4 x 7 H2O	
50	ml	**"Metals 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.


## "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	


## Trace element solution

1.5	g	Na-Nitrilotriacetat	
0.5	g	MnSO4 x H2O	
0.1	g	Fe SO4 x 7 H2O	
0.1	g	Co(NO3)2 x 6 H2O	
0.1	g	ZnCl2	
0.05	g	NiCl2 x 6 H2O	
0.05	g	H2SeO3	
0.01	g	CuSO4 x 5 H2O	
0.01	g	AlK(SO4)2 x 12 H2O	
0.01	g	H3BO3	
0.01	g	Na2MoO4 x 2 H2O	
0.01	g	Na2WO4 x 2 H2O	
1000	ml	Double 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	Double distilled water