Difference between revisions of "WC"
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== Description == | == Description == | ||
− | The WC ( | + | The WC ('''W'''right '''C'''hu) 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).<ref>Guillard, R.R.L. and Lorenzen, C.J. (1972) '''Yellow-green algae with chlorophyllide ''c'''''. ''Journal of Phycology'', 8:1, 10–18. DOI: [https://dx.doi.org/10.1111/j.1529-8817.1972.tb03995.x 10.1111/j.1529-8817.1972.tb03995.x]</ref><ref>Makulla, A. (2000) '''Fatty acid composition of ''Scenedesmus obliquus'': Correlation to dilution rates'''. ''Limnologica'', 30:2, 162–168. DOI: [https://dx.doi.org/10.1016/S0075-9511(00)80011-0 10.1016/S0075-9511(00)80011-0]</ref> |
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== Recipe == | == 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. | ||
+ | {| class="wikitable" | ||
+ | !Buffer | ||
+ | ! | ||
+ | |- | ||
+ | |Compound | ||
+ | |TES | ||
+ | |- | ||
+ | |[M] g/mol | ||
+ | | 229.25 | ||
+ | |- | ||
+ | |mg/5 L | ||
+ | |<nowiki> </nowiki>575 | ||
+ | |- | ||
+ | |mg/L | ||
+ | |<nowiki> </nowiki>115 | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | !Stock solution | ||
+ | !1 | ||
+ | !2 | ||
+ | !3 | ||
+ | !4 | ||
+ | !5 | ||
+ | !6 | ||
+ | |- | ||
+ | |Compound | ||
+ | |CaCl<sub>2</sub> · 2 H<sub>2</sub>O | ||
+ | |MgSO<sub>4</sub> · 7 H<sub>2</sub>O | ||
+ | |NaHCO<sub>3</sub> | ||
+ | |K<sub>2</sub>HPO<sub>4</sub> | ||
+ | |NaNO<sub>3</sub> | ||
+ | | NaSiO<sub>3</sub> · 5 H<sub>2</sub>O | ||
+ | |- | ||
+ | |[M] g/mol | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |- | ||
+ | |g/L | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |- | ||
+ | |g/500 mL | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |- | ||
+ | |g/250 mL | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | !Stock solution | ||
+ | !7: Trace metals | ||
+ | ! | ||
+ | ! | ||
+ | ! | ||
+ | ! | ||
+ | ! | ||
+ | ! | ||
+ | ! | ||
+ | |- | ||
+ | |Compound | ||
+ | |<nowiki> </nowiki>Na<sub>2</sub> · EDTA | ||
+ | |<nowiki> </nowiki>FeCl<sub>3</sub> · 6 H<sub>2</sub>O | ||
+ | |<nowiki> </nowiki>CuSO<sub>4</sub> · 5 H<sub>2</sub>O | ||
+ | |<nowiki> </nowiki>ZnSO<sub>4</sub> · 7 H<sub>2</sub>O | ||
+ | |<nowiki> </nowiki>CoCl<sub>2</sub> · 6 H<sub>2</sub>O | ||
+ | |<nowiki> </nowiki>MnCl<sub>2</sub> · 4 H2O | ||
+ | |<nowiki> </nowiki>NaMoO<sub>4</sub> · 2 H<sub>2</sub>O | ||
+ | |<nowiki> </nowiki>H<sub>3</sub>BO<sub>3</sub> | ||
+ | |- | ||
+ | |[M] g/mol | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |- | ||
+ | |g/L | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |- | ||
+ | |g/500 mL | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |- | ||
+ | |g/250 mL | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | !Stock solution | ||
+ | !8: Vitamins | ||
+ | ! | ||
+ | ! | ||
+ | |- | ||
+ | |Compound | ||
+ | |<nowiki> </nowiki>Thiamin · HCl | ||
+ | |<nowiki> </nowiki>Biotin | ||
+ | |<nowiki> </nowiki>B<sub>12</sub> | ||
+ | |- | ||
+ | |[M] g/mol | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |- | ||
+ | |g/L | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |- | ||
+ | |g/500 mL | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |- | ||
+ | |g/250 mL | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |} | ||
+ | Prepare stock solutions, trace metals, and vitamins in Type I purified water. Stock solution 4 (K<sub>2</sub>HPO<sub>4</sub>) should not be kept in in a polyethylene (PE) bottle and stock solution 6 (NaSiO<sub>3</sub> · 5 H<sub>2</sub>O) should be kept in a polystyrene (PS) bottle instead of a quartz glass bottle.<ref>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: [https://dx.doi.org/10.4319/lo.1963.8.2.0152 10.4319/lo.1963.8.2.0152]</ref><ref>Harrison, P.J. & Berges, J.A. (2005) '''Marine culture media'''. In: ''Algal culturing techniques'' (ed. R.A. Andersen), 1<sup>st</sup> edition, 596 p. Elsevier Academic Press; Burlington, MA, USA.</ref> For vitamins, prepare 1 mL aliquots and keep at -20 °C; thaw to use. | ||
== References == | == References == |
Latest revision as of 16:10, 19 October 2016
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 | ||||||
g/L | ||||||
g/500 mL | ||||||
g/250 mL |
Stock solution | 7: Trace metals | |||||||
---|---|---|---|---|---|---|---|---|
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 | ||||||||
g/L | ||||||||
g/500 mL | ||||||||
g/250 mL |
Stock solution | 8: Vitamins | ||
---|---|---|---|
Compound | Thiamin · HCl | Biotin | B12 |
[M] g/mol | |||
g/L | |||
g/500 mL | |||
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.