TITLE: PCC559 medium
AUTHORS: Roscoff Culture Collection

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span>Pasteur Collection medium for cyanobacteria (</span><span style = "font-style:italic;">Prochlorococcus</span><span>, </span><span style = "font-style:italic;">Synechococcus</span><span>).</span></div><div class = "text-block">Not used by the RCC.</div></div>

[STEPS]
?. [Medium composition]
Under hood, add these nutriments autoclaved before (excepted vitamin):     Quantity
   Compound
     1 L
   Turks Island Salts 1X
     4 mL
   Ferric chloride hexahydrate/EDTA solution
     1mL
   Trace metal for Prochlorococcus medium
     1 mL
   Na-PO4 solution (50 mM, pH 7.5)
     4 mL
   Ammonium sulfate solution (100 mM)
     2 mL
   Sodium hydrogenocarbonate solution (1 M)
     1 mL
   Vitamin B12 (Cyanocobalamin)
     Filter the medium on 0,2microns  AB1Quantity
Compound
21 L
Turks Island Salts 1X
34 mL
Ferric chloride hexahydrate/EDTA solution
41mL
Trace metal for Prochlorococcus medium
51 mL
Na-PO4 solution (50 mM, pH 7.5)
64 mL
Ammonium sulfate solution (100 mM)
72 mL
Sodium hydrogenocarbonate solution (1 M)
81 mL
Vitamin B12 (Cyanocobalamin)
AB1Quantity
Compound
21 L
Turks Island Salts 1X
34 mL
Ferric chloride hexahydrate/EDTA solution
41mL
Trace metal for Prochlorococcus medium
51 mL
Na-PO4 solution (50 mM, pH 7.5)
64 mL
Ammonium sulfate solution (100 mM)
72 mL
Sodium hydrogenocarbonate solution (1 M)
81 mL
Vitamin B12 (Cyanocobalamin)
?. [Turks Island Salts 1X]
Dissolve these salts in the volume of water indicated    Quantity
   Compound
   Volume of dissolution
     28 g
   Sodium chloride (NaCl)
   450 mL
     670 mg
   Potassium chloride (KCl)
   50 mL
     5,5 g
   Magnesium chloride hexahydrate (MgCl2-6H2O)
   100 mL
     6,9 g
   Magnesium sulfate heptahydrate (MgSO4-7H2O)
   150 mL
      1,45 g
   Calcium chloride dihydrate (CaCl2-2H2O)
    100 mL
     Mix the solutions in the order indicatedComplete final volume to 1L of distilled waterAutoclaveStore in refrigerator  ABC1Quantity
Compound
Volume of dissolution
228 g
Sodium chloride (NaCl)
450 mL
3670 mg
Potassium chloride (KCl)
50 mL
45,5 g
Magnesium chloride hexahydrate (MgCl2-6H2O)
100 mL
56,9 g
Magnesium sulfate heptahydrate (MgSO4-7H2O)
150 mL
6&nbsp;1,45 g
Calcium chloride dihydrate (CaCl2-2H2O)
&nbsp;100 mL
Mix the solutions in the order indicatedComplete final volume to 1L of distilled waterAutoclaveStore in refrigerator
ABC1Quantity
Compound
Volume of dissolution
228 g
Sodium chloride (NaCl)
450 mL
3670 mg
Potassium chloride (KCl)
50 mL
45,5 g
Magnesium chloride hexahydrate (MgCl2-6H2O)
100 mL
56,9 g
Magnesium sulfate heptahydrate (MgSO4-7H2O)
150 mL
6&nbsp;1,45 g
Calcium chloride dihydrate (CaCl2-2H2O)
&nbsp;100 mL
?. [Ferric chloride hexahydrate/EDTA solution]
To 10mL of HCl 0,1N, add gradually 270mg of Ferric chloride hexahydrate (FeCl3-6H2O)To 10mL of NaOH 0,1N, add gradually 372mg of Titriplex III dihydrate (EDTA-Na2)Mix both solutionsComplete final volume to 500mL of sterile waterStore in refrigerator
?. [Trace metal for Prochlorococcus medium]
Dissolve these salts in the volume of water indicated :    Quantity
   Compound
   Volume of dissolution
     2.86 g
   Boric acid (H3BO3)
   150 mL
     1.81 g
   Manganese chloride tetrahydrate (MnCl2-4H2O)
   150 mL
     0.222 g
   Zinc sulfate heptahydrate (ZnSO4- 7H2O)
   150 mL
     0.39 g
   Sodium molybdate dihydrate (Na2MoO4-2H2O)
   300 mL
      
 0.049 g
   Cobalt(II) nitrate hexahydrate (Co(NO3)2-6H2O)
    
 150 mL
     Mix the solutions in the order indicatedComplete final volume to 1L of sterile waterStore 6 months in refrigerator  ABC1Quantity
Compound
Volume of dissolution
22.86 g
Boric acid (H3BO3)
150 mL
31.81 g
Manganese chloride tetrahydrate (MnCl2-4H2O)
150 mL
40.222 g
Zinc sulfate heptahydrate (ZnSO4- 7H2O)
150 mL
50.39 g
Sodium molybdate dihydrate (Na2MoO4-2H2O)
300 mL
6&nbsp;
0.049 g
Cobalt(II) nitrate hexahydrate (Co(NO3)2-6H2O)
&nbsp;
150 mL
Mix the solutions in the order indicatedComplete final volume to 1L of sterile waterStore 6 months in refrigerator
ABC1Quantity
Compound
Volume of dissolution
22.86 g
Boric acid (H3BO3)
150 mL
31.81 g
Manganese chloride tetrahydrate (MnCl2-4H2O)
150 mL
40.222 g
Zinc sulfate heptahydrate (ZnSO4- 7H2O)
150 mL
50.39 g
Sodium molybdate dihydrate (Na2MoO4-2H2O)
300 mL
6&nbsp;
0.049 g
Cobalt(II) nitrate hexahydrate (Co(NO3)2-6H2O)
&nbsp;
150 mL
Attention: dilute 10 times with sterile water and filter on 0.2 microns before use
?. [Other solutions]
Na-PO4  solution (50 mM, pH 7.5)Prepare two solutions :  Dissolve 3,45g of monosodium dihydrogen phosphate (NaH2PO4) in 50mL of water
 Dissolve 4,45g of di-sodium hydrogenophosphate dihydrate (Na2HPo4-2H2O) in 50mL of water                                                           
  Make an equimolar mixture of this two solutions and adjust the pH at 7,5Store in refrigeratorAmmonium sulfate solution (100 mM)To 250mL of distilled water, add 3,3g of ammonium sulfate ((NH4)2SO4)Autoclave the solutionStore in refrigeratorSodium hydrogenocarbonate solution (1 M)To 300mL of distilled water, add 25,2g of sodium hydrogenocarbonate (NaHCO3)Autoclave the solutionStore in refrigeratorVitamin B12 (Cyanocobalamin)To 20mL of distilled water, add 20mg of vitamin B12Filter on 0,2 microns (=stock solution at 1mg/mL))Store in freezerThe working solution (10µg/mL) is prepared by aseptical dilution of the vitamin B12 stock solution with sterile waterAmmonium sulfate solution (100 mM)To 250mL of distilled water, add 3,3g of ammonium sulfate ((NH4)2SO4)Autoclave the solutionStore in refrigeratorSodium hydrogenocarbonate solution (1 M)To 300mL of distilled water, add 25,2g of sodium hydrogenocarbonate (NaHCO3)Autoclave the solutionStore in refrigeratorVitamin B12 (Cyanocobalamin)To 20mL of distilled water, add 20mg of vitamin B12Filter on 0,2 microns (=stock solution at 1mg/mL))Store in freezerThe working solution (10µg/mL) is prepared by aseptical dilution of the vitamin B12 stock solution with sterile waterSodium hydrogenocarbonate solution (1 M)To 300mL of distilled water, add 25,2g of sodium hydrogenocarbonate (NaHCO3)Autoclave the solutionStore in refrigeratorVitamin B12 (Cyanocobalamin)To 20mL of distilled water, add 20mg of vitamin B12Filter on 0,2 microns (=stock solution at 1mg/mL))Store in freezerThe working solution (10µg/mL) is prepared by aseptical dilution of the vitamin B12 stock solution with sterile waterVitamin B12 (Cyanocobalamin)To 20mL of distilled water, add 20mg of vitamin B12Filter on 0,2 microns (=stock solution at 1mg/mL))Store in freezerThe working solution (10µg/mL) is prepared by aseptical dilution of the vitamin B12 stock solution with sterile water