Artificial seawater medium
Shaaijer, l.kop, h.koch, Sluecker
Abstract
This artificial seawater medium was adapted from the Synthetic Crenarchaeota Medium described in Könneke et al. 2005 (10.1038/nature03911).
Steps
Prepare the Thauer's vitamin solution and trace element solutions according to the recipes below. Aliquot for future use.
SCMU medium
Dissolve in 900 ml demineralized water:
| A | B |
|---|---|
| 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:
| A | B |
|---|---|
| 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
Dissolve in 1l demineralized water:
| A | B |
|---|---|
| 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)
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)
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:
| A | B |
|---|---|
| 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.
