Difference between revisions of "WC"

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(Created page with " == 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, and the mos...")
 
 
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== Description ==
 
== Description ==
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The WC (Wright Chu) medium is a commonly used fully-defined medium for culturing of limnic algae. It was first described by Guillard & Lorenzen, and the most-commonly used recipe includes modifications as reported by Makulla.
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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 ==
  
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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.
  
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{| class="wikitable"
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!Buffer
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!
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|-
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|Compound
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|TES
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|-
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|[M] g/mol
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|  229.25
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|-
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|mg/5 L
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|<nowiki>  </nowiki>575
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|-
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|mg/L
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|<nowiki>  </nowiki>115
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|}
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{| class="wikitable"
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!Stock solution
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!1
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!2
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!3
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!4
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!5
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!6
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|-
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|Compound
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|CaCl<sub>2</sub> · 2 H<sub>2</sub>O
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|MgSO<sub>4</sub> · 7 H<sub>2</sub>O
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|NaHCO<sub>3</sub>
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|K<sub>2</sub>HPO<sub>4</sub>
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|NaNO<sub>3</sub>
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|  NaSiO<sub>3</sub> · 5 H<sub>2</sub>O
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|-
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|[M] g/mol
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|
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|
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|
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|
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|
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|
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|-
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|g/L
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|
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|
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|
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|
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|
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|
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|-
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|g/500 mL
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|
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|
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|
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|
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|
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|
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|-
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|g/250 mL
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|
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|
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|
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|
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|
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|
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|}
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{| class="wikitable"
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!Stock solution
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!7: Trace metals
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!
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!
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!
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!
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!
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!
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!
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|-
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|Compound
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|<nowiki>  </nowiki>Na<sub>2</sub> · EDTA
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|<nowiki>  </nowiki>FeCl<sub>3</sub> · 6 H<sub>2</sub>O
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|<nowiki>  </nowiki>CuSO<sub>4</sub> · 5 H<sub>2</sub>O
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|<nowiki>  </nowiki>ZnSO<sub>4</sub> · 7 H<sub>2</sub>O
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|<nowiki>  </nowiki>CoCl<sub>2</sub> · 6 H<sub>2</sub>O
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|<nowiki>  </nowiki>MnCl<sub>2</sub> · 4 H2O
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|<nowiki>  </nowiki>NaMoO<sub>4</sub> · 2 H<sub>2</sub>O
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|<nowiki>  </nowiki>H<sub>3</sub>BO<sub>3</sub>
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|-
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|[M] g/mol
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|
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|
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|
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|
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|
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|
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|
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|
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|-
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|g/L
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|
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|
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|
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|
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|
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|
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|
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|
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|-
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|g/500 mL
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|
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|
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|
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|
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|
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|
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|
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|
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|-
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|g/250 mL
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|
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|
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|
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|
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|}
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{| class="wikitable"
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!Stock solution
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!8: Vitamins
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!
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!
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|-
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|Compound
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|<nowiki>  </nowiki>Thiamin · HCl
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|<nowiki>  </nowiki>Biotin
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|<nowiki>  </nowiki>B<sub>12</sub>
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|-
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|[M] g/mol
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|
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|
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|
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|-
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|g/L
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|
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|
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|
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|-
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|g/500 mL
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|
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|
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|
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|-
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|g/250 mL
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|
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|}
  
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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

  1. ↑ 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
  2. ↑ 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
  3. ↑ 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
  4. ↑ 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.
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