User:Sarahfiorini
Ians Cocco Medium (natural seawater base) – version 10/06 ICM (-Tris)-NW
Solution Component FW 1° Stock (g/l)
Working solution (g/l)
Final concentration
Major nutrients
1*
NaNO3
84.99
13.5984
1ml/l⎝
160µM
2* NH4Cl 53.49
0.535
1ml/l⎝
10 µM
3*
KH2PO4
136.09
1.3609
1ml/l⎝
10 µM
4*
(Na)FeEDTA
367.1
8.6
1ml/l⎝
23.4µM
5*
Trace metals
Na2EDTA.2H2O 372.2
37.22
1ml/l⎝
100µM
ZnSO4.7H2O 287.5 11.5 1ml/l⎝
40nM
CoSO4.7H2O 281.1 7.0275 1ml/l⎝
25nM
MnCl2.4H2O 197.9 89.055 1ml/l⎝
450nM
H2SeO3 129.0 0.645 1ml/l⎝
5nM
NiCl2.6H20 149.6 0.745 1ml/l⎝
3.135nM
6**
Vitamins
Vit B12 (cyanocobalamin) 1355.37 0.5 1ml/l⎝ 1ml
1ml/l⎝ 0.37nM
Biotin 244.31 0.005 1ml/l⎝ 1ml
2nM
Thiamine.HCl 337.27
0.1
300nM
- filter sterilize (0.2µm syringe filter), store in fridge (4°C)
- filter sterilize (0.2µm syringe filter), store in freezer (-20°C)
NB: Working Stock -> medium = 1ml/l
1° stocks : 50ml is enough to eventually make 50 000 litres of medium (i.e. you won’t need more than this)
PROTOCOL POUR PREPARER LE MILIEU DE CULTURE
1)Prendre 1 ml de chaque working solution de Major Nutrients (NaNO3, NH4Cl, KH2PO4, NaFeEDTA), les filtrer et les mettre dans un litre d' eau de mer stérile (A).
2)Prendre 37,22 g Na2EDTA.2H et le mettre dans 1 litre de MQ (1). Ajouter à ça 1 ml de chaque stock solution de metaux en trace ( ZnSO4.7H2O, CoSO4.7H2O, MnCl4.4H2O, H2SeO3, NiCl2.6H2O)→ la working solution des metaux en trace est prète (B).
3)Mettre 0.1 g de Thiamine.HCl dans 1L de MQ (2). Ajouter à ça 1 ml de chaque solution stock de vitamines ( B12 et Biotin)→la working solution des vitamines est prète (C).
4)Ajouter 1 ml de B et 1 ml de C aprés les avoir filtrées à A et le milieu est pret pour etre utilisé.
NATURAL SEAWATER MEDIUM PROTOCOL
1)Take 1 ml of every Major Nutrients' working solution (NaNO3, NH4Cl, KH2PO4, NaFeEDTA), filter sterilize and add to 1L sterile seawater (A).
2)Put 37, 22 g Na2EDTA.2H in 1L of MQ (1). Add to this 1 ml of every Trace metals' stock solutions ( ZnSO4.7H2O, CoSO4.7H2O, MnCl4.4H2O, H2SeO3, NiCl2.6H2O)→Trace metals' working solution is now ready (B).
3)Put 0.1 g Thiamine.HCl in 1L of MQ. Add to this 1ml of every Vitamins' stock solution (B12 and Biotin)→Vitamins' working solution is now ready (C).
4) Add to A 1ml of B and C after sterilized and natural medium is ready to use.
