Apr 25, 2024
Provasoli Enriched Seawater (PES) medium solution
- 1School of Biological Sciences, University of Aberdeen, Aberdeen AB24 3FX, United Kingdom
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), 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
LINK
Protocol materials
MilliQ water
Titriplex® III solutionMerck MilliporeSigma (Sigma-Aldrich)
Boric acidFisher ScientificCatalog #BP1681
Manganese sulfate, monohydrateBio Basic Inc.Catalog #MB0334.SIZE.500g
Zinc sulfate heptahydrateMerck MilliporeSigma (Sigma-Aldrich)Catalog #204986
Cobalt(II) sulfate heptahydrateMerck MilliporeSigma (Sigma-Aldrich)
Iron(III) chloride hexahydrateMerck MilliporeSigma (Sigma-Aldrich)Catalog #44944
Ammonium iron(II) sulfate hexahydrateBio Basic Inc.Catalog #AB0065.SIZE.500g
HEPES Sodium saltMerck MilliporeSigma (Sigma-Aldrich)Catalog #H7006
Vitamin B12Fisher ScientificCatalog #68-19-9
Thiamine HClP212121
BiotinP212121
Sodium nitrateMerck MilliporeSigma (Sigma-Aldrich)Catalog #S5506
Sodium glycerophosphate hydrateMerck MilliporeSigma (Sigma-Aldrich)
Potassium iodide
Before start
It is important to follow the order of chemicals added as shown in the protocol.
P II solution
5000 mL MilliQ water
Add 5 g Titriplex® III solutionMerck MilliporeSigma (Sigma-Aldrich)
Add 5.7 g Boric acidFisher ScientificCatalog #BP1681
Add 0.62 g Manganese sulfate, monohydrateBio Basic Inc.Catalog #MB0334.SIZE.500g
Add 0.11 g Zinc sulfate heptahydrateMerck MilliporeSigma (Sigma-Aldrich)Catalog #204986
Add 0.024 g Cobalt(II) sulfate heptahydrateMerck MilliporeSigma (Sigma-Aldrich)
Add 0.245 g Iron(III) chloride hexahydrateMerck MilliporeSigma (Sigma-Aldrich)Catalog #44944
Fe-solution
5000 mL MilliQ water
Add 3.3 g Titriplex® III solutionMerck MilliporeSigma (Sigma-Aldrich)
Add 3.51 g Ammonium iron(II) sulfate hexahydrateBio Basic Inc.Catalog #AB0065.SIZE.500g
Note
Solution equals 0.1 mg Fe2+ x ml-1
Primary solution
2h
6000 mL MilliQ water
Add 23.4 g HEPES Sodium saltMerck MilliporeSigma (Sigma-Aldrich)Catalog #H7006 (or 30 g Tris buffer)
Add 1500 mL P II-solution
Add 1500 mL Fe-solution
Add 21 g Sodium nitrateMerck MilliporeSigma (Sigma-Aldrich)Catalog #S5506
Add 2.12 g Sodium glycerophosphate hydrateMerck MilliporeSigma (Sigma-Aldrich)
Add 16 mg Potassium iodide
Pasteurise for 02:00:00 at 99 °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 60 °C ).
30 mL Vitamin B12Fisher ScientificCatalog #68-19-9 (10 mg per 500 ml MilliQ water )
30 mL Thiamine HClP212121 (500 mg per 500 ml MilliQ water )
30 mL BiotinP212121 (5 mg per 500 ml MilliQ water )
Storage
Store PES in Duran bottles in a cold and dark setting at 10 °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 200 mL PES to 10 L sterile seawater.
Citations
L. Provasoli. Media and prospects for the cultivation of marine algae
https://cir.nii.ac.jp/crid/1572824499547489536