TITLE: PCR S11 - Red Sea medium
AUTHORS: Roscoff Culture Collection

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span>Medium to grow cyanobacteria, in particular </span><span style = "font-style:italic;">Prochlorococcus </span><span>and </span><span style = "font-style:italic;">Synechococcus</span><span></span></div><div class = "text-block"><span style = "font-weight:bold;">Reference</span></div><div class = "text-block"><span>Rippka, R., Coursin, T., Hess, W., Lichtle, C., Scanlan, D.J., Palinska, K.A., Iteman, I. et al. 2000. </span><span style = "font-style:italic;">Prochlorococcus marinus</span><span> Chisholm et al. 1992 subsp. </span><span style = "font-style:italic;">pastoris</span><span> subsp. nov. strain PCC 9511, the first axenic chlorophyll a2/b2-containing cyanobacterium (Oxyphotobacteria). Int. J. Syst. Evol. Microbiol. 50:1833–47. </span></div></div>

[STEPS]
?. [Prepare medium]
We generally prepare two or three 10L carboys at a timeTo 1 L of H2O, add 33.33g of Red Sea Salt Dissolve by shaking (20 min on agitator)Heat seawater during 20min at 100°CTo 1 L of H2O, add 33.33g of Red Sea SaltDissolve by shaking (20 min on agitator)Heat seawater during 20min at 100°C
?. Add to seawater under laminar flow hood the following nutriments that have been autoclaved (except for vitamin) In Roscoff, we generally also add 1 mL NaNO3 1M for Synechococcus (useless for Prochlorococcus but it does not affect the growth)   ABC1Quantity
Compound
Final concentration
21.0 mL
Hepes-NaOH 1M (ph 7,5) -&nbsp;See receipe above
1mM
31.0 mL
Na2-EDTA/FeCl3&nbsp;-&nbsp;See receipe&nbsp;above
2µMa&nbsp;
41.0 mL
Sodium Phosphate (NaPO4) 50mM (pH 7,5)&nbsp;-&nbsp;See receipe&nbsp;above
50µM
51.0 mL
Ammonium Sulfate 400mM (NH4)2-SO4
400µM
60.1 mLb
Trace metals “Gaffron+Se” -&nbsp;See receipe&nbsp;above
&nbsp;
7&nbsp;
0.1 mL
Cyanocobalamin 10mg/L (Vit. B12)
1µg/Lc
ABC1Quantity
Compound
Final concentration
21.0 mL
Hepes-NaOH 1M (ph 7,5) -&nbsp;See receipe above
1mM
31.0 mL
Na2-EDTA/FeCl3&nbsp;-&nbsp;See receipe&nbsp;above
2µMa&nbsp;
41.0 mL
Sodium Phosphate (NaPO4) 50mM (pH 7,5)&nbsp;-&nbsp;See receipe&nbsp;above
50µM
51.0 mL
Ammonium Sulfate 400mM (NH4)2-SO4
400µM
60.1 mLb
Trace metals “Gaffron+Se” -&nbsp;See receipe&nbsp;above
&nbsp;
7&nbsp;
0.1 mL
Cyanocobalamin 10mg/L (Vit. B12)
1µg/Lc
a - The original recipe is 8 µM (Rippka et al 2000 IJSEM 50, 1833–1847). But it works fine for Prochlorococcus...b -  If one adds 0.1 mL par L, the final concentration of Gaffron in the PCR-S11 medium is actually twice more than in the original PCR-S11 recipe.c - 10 times less than in the original receipe. Again this seems sufficient...
?. Filter the medium on 0.2 micron filter
?. [Prepare solutions]
Hepes-NaOH 1MTo 250 mL of H20, add gradually 119.15g of Hepes.Adjust pH at 7.5 and complete the volume at 500 mL.Store in refrigerator.
?. Na2-EDTA/FeCl3To 40 mL of HCl 0.1 N, add gradually 1,080 g of FeCl3To 40 mL of NaOH 0.1 N, add gradually 1,488 g of Na2-EDTAMix both solutionsComplete final volume to 2 L of sterile waterStore in refrigerator
?. Sodium PhosphatePrepare two solutions :  Monosodium dihydrogen phosphate (NaH2PO4) at 50 mM (6 g in 1 L)Disodium hydrogen phosphate (Na2HPo4) at 50 mM (3.55 g in 500 mL)Make an equimolar mixture of these two solutions and adjust the pH at 7,5
?. To 500 mL of H20, add gradually elements indicated in the table below. Complement the volume to 1L. Store in refrigerator.  ABCDE1Quantity (mg/L)
Merck ref.
&nbsp;
Compound
Concentration in stock solution
(µM)Final&nbsp;&nbsp; concentration in medium
(nM)2186
165
Boric acid (H3BO3)
3008.25300.823101
596
Manganese (II) Sulfate Monohydrate (MnSO4.H2O)
597.5659.7641.98
6673
Sodium Tungstate dihydrate (Na2WO4.2H20)
60.655.16b
1182
Ammonium molybdate tetrahydrate ((NH4)6MO7O24.4H2O)
4.180.4267.14
4905
Potassium bromide (KBr)
60674.98
5043
Potassium iodide (KI)
303817.25
8883
Zinc sulfate heptahydrate (ZnSO4.7H2O)
60699.25
2019
Cadium Nitrate (Cd(NO3)2.4H2O)
303108.76
2554
Cobalt (II) Nitrate (Co(NO3)2.6H2O)
303117.5
2790
Copper (II) Sulfate (CuSO4.5H2O)
303127.1
6717
Nickel Chloride (NiCl2.6H2O)
303132.4
2481
Chromium (III) Nitrate (Cr(NO3)3.9H2O)
60.6141.5c
8503
Vanadyl Sulfate Pentahydrate (VOSO4.5H2O)5.930.591528.4
1047
Aluminium Potassium Sulfate (KAl(SO4)2.12H2O)
59.875.99163.3
800653
Selenium (IV) Oxyde (SeO2)
29.742.97
ABCDE1Quantity (mg/L)
Merck ref.
&nbsp;
Compound
Concentration in stock solution
(µM)Final&nbsp;&nbsp; concentration in medium
(nM)2186
165
Boric acid (H3BO3)
3008.25300.823101
596
Manganese (II) Sulfate Monohydrate (MnSO4.H2O)
597.5659.7641.98
6673
Sodium Tungstate dihydrate (Na2WO4.2H20)
60.655.16b
1182
Ammonium molybdate tetrahydrate ((NH4)6MO7O24.4H2O)
4.180.4267.14
4905
Potassium bromide (KBr)
60674.98
5043
Potassium iodide (KI)
303817.25
8883
Zinc sulfate heptahydrate (ZnSO4.7H2O)
60699.25
2019
Cadium Nitrate (Cd(NO3)2.4H2O)
303108.76
2554
Cobalt (II) Nitrate (Co(NO3)2.6H2O)
303117.5
2790
Copper (II) Sulfate (CuSO4.5H2O)
303127.1
6717
Nickel Chloride (NiCl2.6H2O)
303132.4
2481
Chromium (III) Nitrate (Cr(NO3)3.9H2O)
60.6141.5c
8503
Vanadyl Sulfate Pentahydrate (VOSO4.5H2O)5.930.591528.4
1047
Aluminium Potassium Sulfate (KAl(SO4)2.12H2O)
59.875.99163.3
800653
Selenium (IV) Oxyde (SeO2)
29.742.97
a -The original receipe uses 300 nM H3BO3 finalb - 4.94 in the original receipec - 1.52 in the original receipe