Apr 25, 2024

Public workspaceProvasoli Enriched Seawater (PES) medium solution

DOI
dx.doi.org/10.17504/protocols.io.rm7vzyyo5lx1/v1
  • 1School of Biological Sciences, University of Aberdeen, Aberdeen AB24 3FX, United Kingdom
Anton Kuech
Open access
DOI: 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 07, 2022
Last Modified: April 25, 2024
Protocol Integer ID: 57898
Keywords: Macroalgae, Seaweed cultivation, Enriched seawater medium, Cultivation medium, Provasoli enriched seawater (PES)
Funders Acknowledgement:
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.
LINK
https://cir.nii.ac.jp/crid/1572824499547489536

Protocol materials
ReagentMilliQ water
In 6 steps
ReagentTitriplex® III solutionMerck MilliporeSigma (Sigma-Aldrich)
In 2 steps
ReagentZinc sulfate heptahydrateMerck MilliporeSigma (Sigma-Aldrich)Catalog #204986
Step 1.4
ReagentBiotinP212121
Step 4.3
ReagentPotassium iodide
Step 3.6
ReagentVitamin B12Fisher ScientificCatalog #68-19-9
Step 4.1
ReagentSodium glycerophosphate hydrateMerck MilliporeSigma (Sigma-Aldrich)
Step 3.5
ReagentBoric acidFisher ScientificCatalog #BP1681
Step 1.2
ReagentManganese sulfate, monohydrateBio Basic Inc.Catalog #MB0334.SIZE.500g
Step 1.3
ReagentCobalt(II) sulfate heptahydrateMerck MilliporeSigma (Sigma-Aldrich)
Step 1.5
ReagentIron(III) chloride hexahydrateMerck MilliporeSigma (Sigma-Aldrich)Catalog #44944
Step 1.6
ReagentHEPES Sodium saltMerck MilliporeSigma (Sigma-Aldrich)Catalog #H7006
Step 3.1
ReagentSodium nitrateMerck MilliporeSigma (Sigma-Aldrich)Catalog #S5506
Step 3.4
ReagentAmmonium iron(II) sulfate hexahydrateBio Basic Inc.Catalog #AB0065.SIZE.500g
Step 2.2
ReagentThiamine HClP212121
Step 4.2
Before start
It is important to follow the order of chemicals added as shown in the protocol.
P II solution
P II solution
Amount5000 mL ReagentMilliQ waterSigma Aldrich

Add Amount5 g ReagentTitriplex® III solutionSigma Aldrich
Add Amount5.7 g ReagentBoric acidSigma AldrichCatalog #BP1681

Add Amount0.62 g ReagentManganese sulfate, monohydrateSigma AldrichCatalog #MB0334.SIZE.500g

Add Amount0.11 g ReagentZinc sulfate heptahydrateSigma AldrichCatalog #204986

Add Amount0.024 g ReagentCobalt(II) sulfate heptahydrateSigma Aldrich

Add Amount0.245 g ReagentIron(III) chloride hexahydrateSigma AldrichCatalog #44944

Fe-solution
Fe-solution
Amount5000 mL ReagentMilliQ waterSigma Aldrich

Add Amount3.3 g ReagentTitriplex® III solutionSigma Aldrich

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



Primary solution
Primary solution
2h
Amount6000 mL ReagentMilliQ waterSigma Aldrich
Add Amount23.4 g ReagentHEPES Sodium saltSigma AldrichCatalog #H7006 (or Amount30 g Tris buffer)
Add Amount1500 mL P II-solution Go togo to step #1
Add Amount1500 mL Fe-solution Go togo to step #2
Add Amount21 g ReagentSodium nitrateSigma AldrichCatalog #S5506
Add Amount2.12 g ReagentSodium glycerophosphate hydrateMerck MilliporeSigma (Sigma-Aldrich)

Add Amount16 mg ReagentPotassium iodideSigma Aldrich
Pasteurise for Duration02:00:00 at Temperature99 °C
2h
Vitamins
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 B12Sigma AldrichCatalog #68-19-9 (10 mg per 500 ml ReagentMilliQ waterSigma Aldrich )
Amount30 mL ReagentThiamine HClSigma Aldrich (500 mg per 500 ml ReagentMilliQ waterSigma Aldrich )
Amount30 mL ReagentBiotinSigma Aldrich (5 mg per 500 ml ReagentMilliQ waterSigma Aldrich )
Storage
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
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