TITLE: K and K+Si medium
AUTHORS: Roscoff Culture Collection

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Medium to grow most phytoplankton species in small eukaryotes (Mamiellophyceae, Pelagophyceae etc...).</div><div class = "text-block">When using for diatoms and coccolithophorids add silica (Si).</div><div class = "text-block">Reference</div><div class = "text-block">Keller, M.D., Selvin, R.C., Claus, W. & Guillard, R.R.L. 1987. Media for the culture of oceanic ultraphytoplankton. J. Phycol. 23:633–8.</div></div>

[STEPS]
?. Filter 1L of old seawater of at least two months on prefilter and filter 0,2 microns
?. Heat seawater during 20min at 100°C
?. Under hood, to seawater, add these nutriments that have been autoclaved (excepted vitamin) :  ABC1Quantity
Compound
Stock Solution
21.0 mL
Sodium Nitrate (NaNO3)
75.0 g/L of H2O
31.0 mL
Ammonium Chloride (NH4Cl)
2.68 g/L of H2O
41.0 mL
Na2-Glycérophosphate (C3H7O6PNa2, 5 à 6 H2O)
3.06 g/L of H2O
51.0 mL
TRIS-Base (pH7.2)
121.1g/L of H2O
61.0 mL
K Trace Metal Solution
(see recipe below)
70.1 mL
f/2 Vitamin Solution
(see recipe below)
8&nbsp;&nbsp;&nbsp;9For K+Si&nbsp;
Add
&nbsp;101.0 mLSodium Metasilicate Nonahydrate (Na2SiO3.9H2O)30 g/L of H2O
ABC1Quantity
Compound
Stock Solution
21.0 mL
Sodium Nitrate (NaNO3)
75.0 g/L of H2O
31.0 mL
Ammonium Chloride (NH4Cl)
2.68 g/L of H2O
41.0 mL
Na2-Glycérophosphate (C3H7O6PNa2, 5 à 6 H2O)
3.06 g/L of H2O
51.0 mL
TRIS-Base (pH7.2)
121.1g/L of H2O
61.0 mL
K Trace Metal Solution
(see recipe below)
70.1 mL
f/2 Vitamin Solution
(see recipe below)
8&nbsp;&nbsp;&nbsp;9For K+Si&nbsp;
Add
&nbsp;101.0 mLSodium Metasilicate Nonahydrate (Na2SiO3.9H2O)30 g/L of H2O
?. K Trace Metal SolutionTo 900 mL of H2O add :     Quantity
   Compound
   Stock Solution
     41,6g
   EDTA Disodium Salt Dihydrate (Na2EDTA.2H2O)
    
     3,15g
   Hexahydrated ferric chloride (FeCl3.6H2O)
   1,5 cc (liquide)
     1.0 ml
   Sodium Molybdate Dihydrate (Na2MoO4.2H2O)
   6,3 g/L of H2O
     1.0 ml
   Zinc Sulfate Heptahydrate (ZnSO4.7H2O)
   22.0g/L of H2O
     1.0 ml
   Cobalt Chloride Hexahydrate (CoCl2.6H2O)
   10.0g/L of H2O
     1.0 ml
   Manganese (II) chloride, tetrahydrate (MnCl2.4H2O)
   180.0g/L of H2O
     1.0 ml
   Copper(II) sulfate pentahydrate (Cu SO4.5H2O)
   4.9g/L of H2O
     1.0 ml
   Selenous acid (H2SeO3)
   1,29g/L of H2O
     Make final volume up to 1.0L using H2O. Heat to dissolve  ABC1Quantity
Compound
Stock Solution
241,6g
EDTA Disodium Salt Dihydrate (Na2EDTA.2H2O)
&nbsp;
33,15g
Hexahydrated ferric chloride (FeCl3.6H2O)
1,5 cc (liquide)
41.0 ml
Sodium Molybdate Dihydrate (Na2MoO4.2H2O)
6,3 g/L of H2O
51.0 ml
Zinc Sulfate Heptahydrate (ZnSO4.7H2O)
22.0g/L of H2O
61.0 ml
Cobalt Chloride Hexahydrate (CoCl2.6H2O)
10.0g/L of H2O
71.0 ml
Manganese (II) chloride, tetrahydrate (MnCl2.4H2O)
180.0g/L of H2O
81.0 ml
Copper(II) sulfate pentahydrate (Cu SO4.5H2O)
4.9g/L of H2O
91.0 ml
Selenous acid (H2SeO3)
1,29g/L of H2O
ABC1Quantity
Compound
Stock Solution
241,6g
EDTA Disodium Salt Dihydrate (Na2EDTA.2H2O)
&nbsp;
33,15g
Hexahydrated ferric chloride (FeCl3.6H2O)
1,5 cc (liquide)
41.0 ml
Sodium Molybdate Dihydrate (Na2MoO4.2H2O)
6,3 g/L of H2O
51.0 ml
Zinc Sulfate Heptahydrate (ZnSO4.7H2O)
22.0g/L of H2O
61.0 ml
Cobalt Chloride Hexahydrate (CoCl2.6H2O)
10.0g/L of H2O
71.0 ml
Manganese (II) chloride, tetrahydrate (MnCl2.4H2O)
180.0g/L of H2O
81.0 ml
Copper(II) sulfate pentahydrate (Cu SO4.5H2O)
4.9g/L of H2O
91.0 ml
Selenous acid (H2SeO3)
1,29g/L of H2O
?. Filter the medium on 0.2 µm.
?. f/2 Vitamin solutionIn 100mL of distilled water, dissolve 0,05g of biotin (vit. H) and 0,05g of cyanocobalamin (vit. B12) = solution AIn 0,5mL of solution A, add 0,05g of thiamine HCl (vit. B1) and complete final volume to 50mL of distilled waterFilter sterilize on Millipore 0,22µmStore in refrigerator or freezer