Provasoli Enriched Seawater (PES) medium solution

Anton Kuech

Published: 2024-04-25 DOI: 10.17504/protocols.io.rm7vzyyo5lx1/v1

Provasoli enriched seawater (PES)

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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 Media and prospects for the cultivation of marine algae Proceedings of the US-Japan Conference https://cir.nii.ac.jp/crid/1572824499547489536

Before start

It is important to follow the order of chemicals added as shown in the protocol.

Steps

P II solution

5000mL

1.1.

Add 5g

1.2.

Add 5.7g

1.3.

Add 0.62g

1.4.

Add 0.11g

1.5.

Add 0.024g

1.6.

Add 0.245g

Fe-solution

5000mL

2.1.

Add 3.3g

2.2.

Add 3.51g

Note

Solution equals 0.1mg Fe²⁺ x ml^-1

Primary solution

6000mL

3.1.

Add 23.4g (or 30g Tris buffer)

3.2.

Add 1500mL P II-solution

3.3.

Add 1500mL Fe-solution

3.4.

Add 21g

3.5.

Add 2.12g

3.6.

Add 16mg

3.7.

Pasteurise for 2h 0m 0s at 99°C

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 ).

4.1.

30mL (10 mg per 500 ml )

4.2.

30mL (500 mg per 500 ml )

4.3.

30mL (5 mg per 500 ml )

Storage

Store PES in Duran bottles in a cold and dark setting at 10°C .

Equipment

Value	Label
Duran® laboratory bottles, with caps	NAME
Glass bottle	TYPE
Duran	BRAND
Z305197-10EA	SKU
http://sigmaaldrich.com	LINK
500 ml	SPECIFICATIONS

Application

Culturing medium is created by adding 200mL PES to 10L sterile seawater.