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

NB: Working Stock -> medium = 1ml/l
 * filter sterilize (0.2µm syringe filter), store in fridge (4°C)
 * filter sterilize (0.2µm syringe filter), store in freezer (-20°C)

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.