Description
The WC (Wright Chu) medium is a commonly used fully-defined medium for culturing of limnic algae. It was first described by Guillard & Lorenzen (1972), and the most-commonly used recipe includes modifications as reported by Makulla (2000).[1][2]
Recipe
Dissolve TES buffer in Type I purified water (e.g., Milli-Q) using a magnetic stirrer and add 1 mL/L of stock solutions and trace metals in numerical order (1–7). Autoclave appropriately and add 1 mL/L vitamins (8) through a sterile 0.22 μm polystyrene (PES) filter once cooled down to room temperature.
Buffer
|
|
Compound
|
TES
|
[M] g/mol
|
229.25
|
mg/5 L
|
575
|
mg/L
|
115
|
Stock solution
|
1
|
2
|
3
|
4
|
5
|
6
|
Compound
|
CaCl2 · 2 H2O
|
MgSO4 · 7 H2O
|
NaHCO3
|
K2HPO4
|
NaNO3
|
NaSiO3 · 5 H2O
|
[M] g/mol
|
|
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g/L
|
|
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g/500 mL
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|
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g/250 mL
|
|
|
|
|
|
|
Stock solution
|
7: Trace metals
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|
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|
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Compound
|
Na2 · EDTA
|
FeCl3 · 6 H2O
|
CuSO4 · 5 H2O
|
ZnSO4 · 7 H2O
|
CoCl2 · 6 H2O
|
MnCl2 · 4 H2O
|
NaMoO4 · 2 H2O
|
H3BO3
|
[M] g/mol
|
|
|
|
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g/L
|
|
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g/500 mL
|
|
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g/250 mL
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|
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|
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|
Stock solution
|
8: Vitamins
|
|
|
Compound
|
Thiamin · HCl
|
Biotin
|
B12
|
[M] g/mol
|
|
|
|
g/L
|
|
|
|
g/500 mL
|
|
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|
g/250 mL
|
|
|
|
Prepare stock solutions, trace metals, and vitamins in Type I purified water. Stock solution 4 (K2HPO4) should not be kept in in a polyethylene (PE) bottle and stock solution 6 (NaSiO3 · 5 H2O) should be kept in a polystyrene (PS) bottle instead of a quartz glass bottle.[3][4] For vitamins, prepare 1 mL aliquots and keep at -20 °C; thaw to use.
References
- ↑ Guillard, R.R.L. and Lorenzen, C.J. (1972) Yellow-green algae with chlorophyllide c. Journal of Phycology, 8:1, 10–18. DOI: 10.1111/j.1529-8817.1972.tb03995.x
- ↑ Makulla, A. (2000) Fatty acid composition of Scenedesmus obliquus: Correlation to dilution rates. Limnologica, 30:2, 162–168. DOI: 10.1016/S0075-9511(00)80011-0
- ↑ Hassenteufel, W., Jagitsch, R. & Koczy, F.F. (1963) Impregnation of glass surface against sorption of phosphate traces. Limnology and Oceanography, 8:2, 152–156. DOI: 10.4319/lo.1963.8.2.0152
- ↑ Harrison, P.J. & Berges, J.A. (2005) Marine culture media. In: Algal culturing techniques (ed. R.A. Andersen), 1st edition, 596 p. Elsevier Academic Press; Burlington, MA, USA.