Apr 25, 2024

Public workspaceProvasoli Enriched Seawater (PES) medium solution

DOI
https://dx.doi.org/10.17504/protocols.io.rm7vzyyo5lx1/v1
  • Anton Kuech1
  • 1School of Biological Sciences, University of Aberdeen, Aberdeen AB24 3FX, United Kingdom
Anton Kuech
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DOI: https://dx.doi.org/10.17504/protocols.io.rm7vzyyo5lx1/v1
Protocol Citation: Anton Kuech 2024. Provasoli Enriched Seawater (PES) medium solution. protocols.io https://dx.doi.org/10.17504/protocols.io.rm7vzyyo5lx1/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: February 08, 2022
Last Modified: April 25, 2024
Protocol Integer ID: 57898
Keywords: Macroalgae, Seaweed cultivation, Enriched seawater medium, Cultivation medium, Provasoli enriched seawater (PES), provasoli enriched seawater, method by provasoli, medium solution, provasoli, medium solution this protocol, pe, medium in accordance, seawater
Funders Acknowledgements:
Natural Environment Research Council
Grant ID: NE/S007377/1
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Abstract
This protocol outlines the steps required to prepare Provasoli Enriched Seawater (PES) medium in accordance with the method by Provasoli (1966).
Citation
L. Provasoli (1966). Media and prospects for the cultivation of marine algae. Proceedings of the US-Japan Conference.
https://cir.nii.ac.jp/crid/1572824499547489536
LINK

Protocol materials
ReagentMilliQ water
ReagentTitriplex® III solutionMerck MilliporeSigma (Sigma-Aldrich)
ReagentBoric acidFisher ScientificCatalog #BP1681
ReagentManganese sulfate, monohydrateBio Basic Inc.Catalog #MB0334.SIZE.500g
ReagentZinc sulfate heptahydrateMerck MilliporeSigma (Sigma-Aldrich)Catalog #204986
ReagentCobalt(II) sulfate heptahydrateMerck MilliporeSigma (Sigma-Aldrich)
ReagentIron(III) chloride hexahydrateMerck MilliporeSigma (Sigma-Aldrich)Catalog #44944
ReagentAmmonium iron(II) sulfate hexahydrateBio Basic Inc.Catalog #AB0065.SIZE.500g
ReagentHEPES Sodium saltMerck MilliporeSigma (Sigma-Aldrich)Catalog #H7006
ReagentVitamin B12Fisher ScientificCatalog #68-19-9
ReagentThiamine HClP212121
ReagentBiotinP212121
ReagentSodium nitrateMerck MilliporeSigma (Sigma-Aldrich)Catalog #S5506
ReagentSodium glycerophosphate hydrateMerck MilliporeSigma (Sigma-Aldrich)
ReagentPotassium iodide
Before start
It is important to follow the order of chemicals added as shown in the protocol.
P II solution
Amount5000 mL ReagentMilliQ water

Add Amount5 g ReagentTitriplex® III solutionMerck MilliporeSigma (Sigma-Aldrich)
Add Amount5.7 g ReagentBoric acidFisher ScientificCatalog #BP1681

Add Amount0.62 g ReagentManganese sulfate, monohydrateBio Basic Inc.Catalog #MB0334.SIZE.500g

Add Amount0.11 g ReagentZinc sulfate heptahydrateMerck MilliporeSigma (Sigma-Aldrich)Catalog #204986

Add Amount0.024 g ReagentCobalt(II) sulfate heptahydrateMerck MilliporeSigma (Sigma-Aldrich)

Add Amount0.245 g ReagentIron(III) chloride hexahydrateMerck MilliporeSigma (Sigma-Aldrich)Catalog #44944

Fe-solution
Amount5000 mL ReagentMilliQ water

Add Amount3.3 g ReagentTitriplex® III solutionMerck MilliporeSigma (Sigma-Aldrich)

Add Amount3.51 g ReagentAmmonium iron(II) sulfate hexahydrateBio Basic Inc.Catalog #AB0065.SIZE.500g
Note
Solution equals Amount0.1 mg Fe2+ x ml-1



Primary solution
2h
Amount6000 mL ReagentMilliQ water
Add Amount23.4 g ReagentHEPES Sodium saltMerck MilliporeSigma (Sigma-Aldrich)Catalog #H7006 (or Amount30 g Tris buffer)
Add Amount1500 mL P II-solution Go to
Add Amount1500 mL Fe-solution Go to
Add Amount21 g ReagentSodium nitrateMerck MilliporeSigma (Sigma-Aldrich)Catalog #S5506
Add Amount2.12 g ReagentSodium glycerophosphate hydrateMerck MilliporeSigma (Sigma-Aldrich)

Add Amount16 mg ReagentPotassium iodide
Pasteurise for Duration02:00:00 at Temperature99 °C
2h
Vitamins
Vitamins to be added through a sterile single injection with a front filter (0.2 μm) in the cooled off primary solution (maximum temperature of Temperature60 °C ).
Amount30 mL ReagentVitamin B12Fisher ScientificCatalog #68-19-9 (10 mg per 500 ml ReagentMilliQ water )
Amount30 mL ReagentThiamine HClP212121 (500 mg per 500 ml ReagentMilliQ water )
Amount30 mL ReagentBiotinP212121 (5 mg per 500 ml ReagentMilliQ water )
Storage
Store PES in Duran bottles in a cold and dark setting at Temperature10 °C .
Equipment
Duran® laboratory bottles, with caps
NAME
Glass bottle
TYPE
Duran
BRAND
Z305197-10EA
SKU
500 ml
SPECIFICATIONS

Application
Culturing medium is created by adding Amount200 mL PES to Amount10 L sterile seawater.
Citations
L. Provasoli. Media and prospects for the cultivation of marine algae
https://cir.nii.ac.jp/crid/1572824499547489536