Protocol Citation: Shaaijer, l.kop, h.koch, Sluecker 2022. Artificial seawater medium. protocols.io https://dx.doi.org/10.17504/protocols.io.81wgby1zovpk/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: September 12, 2022
Last Modified: December 16, 2022
Protocol Integer ID: 69881
Keywords: Artificial seawater medium, seawater growth medium, synthetic seawater
Funders Acknowledgement:
Netherlands Organisation for Scientific Research
Grant ID: 016.Vidi.189.050
Netherlands Organisation for Scientific Research
Grant ID: VI.Veni.192.086
Dutch Ministry of Education, Culture and Science
Grant ID: 024.002.002 (SIAM)
Abstract
This artificial seawater medium was adapted from the Synthetic Crenarchaeota Medium described in Könneke et al. 2005 (10.1038/nature03911).
Prepare the Thauer's vitamin solution and trace element solutions according to the recipes below. Aliquot for future use.
SCMU medium
SCMU medium
Dissolve in 900 ml demineralized water:
| compound | weight in g (for 900 ml medium) | |
| NaCl | 22 | |
| MgCl2 x 6 H2O | 4.5 | |
| MgSO4 x 7 H2O | 4.5 | |
| CaCl2 x 2 H2O | 0.675 |
Autoclave at 121°C for 20 min at 15 psi.
After cooling to room temperature add (in the flow cabinet) from sterile stocks:
| compound | amount (ml) | |
| 1M KH2PO4 | 0.261 | |
| TES I | 0.45 | |
| TES II | 0.9 | |
| Thauer’s vitamin solution | 0.45 | |
| NaHCO3 | 1.8 |
Adjust the pH to 7.8 using sterile 1M NaHCO3 solution.
Thauer's vitamin solution
Thauer's vitamin solution
Dissolve in 1l demineralized water:
| compound | weight in g (for 1 l) | |
| Biotin | 0.02 | |
| Folic Acid | 0.02 | |
| Pyridoxine HCl | 0.1 | |
| Thiamine HCl | 0.05 | |
| Riboflavin | 0.05 | |
| Nicotinic Acid | 0.05 | |
| DL Pantothenic Acid | 0.05 | |
| p Aminobenzoic Acid | 0.05 | |
| Choline Chloride | 2 | |
| Vitamin B12 | 0.01 |
Adjust to pH 7.0 with KOH.
Filter sterilize (0.2 µm) and store at 4ºC.
Trace Elements Solution I (TES I)
Trace Elements Solution I (TES I)
Dissolve 10 g NTA (Trisodium Nitrilotriacetate) in 500 ml demineralized water.
Adjust to pH 8 using NaOH pellets.
Add 5 g FeSO4 x 7 H2O and dissolve.
Top up with demineralized water to a total volume of 1l.
Filter sterilize (0.2 µm) or autoclave at 121°C for 20 min at 15 psi.
Store in the dark (slightly light sensitive) at RT.
Trace Elements Solution II (TES II)
Trace Elements Solution II (TES II)
Dissolve 5.5 g NTA (Trisodium Nitrilotriacetate) in 500 ml demineralized water.
Adjust to pH 8 using NaOH pellets.
Add the following ingredients and dissolve:
| compound | weight in g (for 1 l) | |
| ZnSO4 x 7 H2O | 0.43 | |
| CoCl2 x 6 H2O | 0.24 | |
| MnCl2 x 4 H2O | 0.99 | |
| CuSO4 x 5 H2O | 0.25 | |
| Na2MoO4 x 2 H2O | 0.22 | |
| NiCl2 x 6 H2O | 0.19 | |
| NaSeO4 | 0.1075 | |
| H3BO3 | 0.14 | |
| CeCl3 x 7 H2O | 0.24 |
Top up with demineralized water to a total volume of 1l.
Filter sterilize (0.2 µm) or autoclave at 121°C for 20 min at 15 psi.
Store in the dark (slightly light sensitive) at RT.