Difference between revisions of "WC"

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== Description ==
 
== Description ==
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<ref>Guillard, R.R.L. and Lorenzen, C.J. (1972) '''Yellow-green algae with chlorophyllide ''c'''''. ''Journal of Phycology'', 8:1, 10–18. DOI: </ref>, and the most-commonly used recipe includes modifications as reported by Makulla<ref>Makulla, A. (2000) '''Fatty acid composition of ''Scenedesmus obliquus'': Correlation to dilution rates'''. ''Limnologica'', 30:2, 162–168. DOI:</ref>.
+
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>
 
== 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.
 
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"
 
{| 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
 +
|
 +
|
 +
|
 +
|
 
|
 
|
 
|
 
|
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|
 
|
 
|-
 
|-
 +
|g/500 mL
 +
|
 +
|
 +
|
 +
|
 
|
 
|
 
|
 
|
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|
 
|
 
|-
 
|-
 +
|g/250 mL
 +
|
 +
|
 +
|
 +
|
 
|
 
|
 
|
 
|
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|
 
|
 
|}
 
|}
{| class="MsoTableGrid"
+
{| class="wikitable"
+
!Stock solution
  | colspan="2" |
+
!8: Vitamins
<nowiki>  </nowiki>'''Buffer'''
+
!
 
+
!
  |
+
|-
<nowiki>  </nowiki>[M] g/mol
+
|Compound
 
+
|<nowiki>  </nowiki>Thiamin · HCl
  |
+
|<nowiki>  </nowiki>Biotin
<nowiki>  </nowiki>mg/5 L
+
|<nowiki>  </nowiki>B<sub>12</sub>
 
+
|-
  |
+
|[M] g/mol
<nowiki>  </nowiki>mg/L
+
|
 
+
|
  |   
+
|
+
|-
|-
+
|g/L
  | 
+
|
 
+
|
  |
+
|
<nowiki>  </nowiki>TES
+
|-
 
+
|g/500 mL
  |
+
|
<nowiki>  </nowiki>229.25
+
|
 
+
|
  |
+
|-
<nowiki>  </nowiki>575
+
|g/250 mL
 
+
|
  |
+
|
<nowiki>  </nowiki>115
+
|
 
 
  |   
 
 
|-
 
  | 
 
 
 
  | 
 
 
 
  | 
 
 
 
  | 
 
 
 
  | 
 
 
 
  |   
 
 
|-
 
  | colspan="2" |
 
<nowiki>  </nowiki>'''Stock
 
<nowiki> </nowiki> solutions'''
 
 
 
  |
 
<nowiki>  </nowiki>[M] g/mol
 
 
 
  |
 
<nowiki>  </nowiki>g/L
 
 
 
  |
 
<nowiki>  </nowiki>g/500 mL
 
 
 
  |
 
<nowiki>  </nowiki>g/250 mL 
 
 
|-
 
  |
 
<nowiki>  </nowiki>'''1'''
 
 
 
  |
 
<nowiki>  </nowiki>CaCl<sub>2</sub> · 2 H<sub>2</sub>O
 
 
 
  |
 
<nowiki>  </nowiki>147.02
 
 
 
  |
 
<nowiki>  </nowiki>36.8
 
 
 
  |
 
<nowiki>  </nowiki>18.4
 
 
 
  |
 
<nowiki>  </nowiki>9.20 
 
 
|-
 
  |
 
<nowiki>  </nowiki>'''2'''
 
 
 
  |
 
<nowiki>  </nowiki>MgSO<sub>4</sub> · 7 H<sub>2</sub>O
 
 
 
  |
 
<nowiki>  </nowiki>246.48
 
 
 
  |
 
<nowiki>  </nowiki>37.0
 
 
 
  |
 
<nowiki>  </nowiki>18.5
 
 
 
  |
 
<nowiki>  </nowiki>9.25 
 
 
|-
 
  |
 
<nowiki>  </nowiki>'''3'''
 
 
 
  |
 
<nowiki>  </nowiki>NaHCO<sub>3</sub>
 
 
 
  |
 
<nowiki>  </nowiki>84.01
 
 
 
  |
 
<nowiki>  </nowiki>12.6
 
 
 
  |
 
<nowiki>  </nowiki>6.3
 
 
 
  |
 
<nowiki>  </nowiki>3.15 
 
 
|-
 
  |
 
<nowiki>  </nowiki>'''4'''
 
 
 
  |
 
<nowiki>  </nowiki>K<sub>2</sub>HPO<sub>4</sub>
 
 
 
  |
 
<nowiki>  </nowiki>174.18
 
 
 
  |
 
<nowiki>  </nowiki>8.7
 
 
 
  |
 
<nowiki>  </nowiki>4.4
 
 
 
  |
 
<nowiki>  </nowiki>2.18 
 
 
|-
 
  |
 
<nowiki>  </nowiki>'''5'''
 
 
 
  |
 
<nowiki>  </nowiki>NaNO<sub>3</sub>
 
 
 
  |
 
<nowiki>  </nowiki>85.01
 
 
 
  |
 
<nowiki>  </nowiki>85.0
 
 
 
  |
 
<nowiki>  </nowiki>42.5
 
 
 
  |
 
<nowiki>  </nowiki>21.25 
 
 
|-
 
  |
 
<nowiki>  </nowiki>'''6'''
 
 
 
  |
 
<nowiki>  </nowiki>NaSiO<sub>3</sub> · 5 H<sub>2</sub>O
 
 
 
  |
 
<nowiki>  </nowiki>212.16
 
 
 
  |
 
<nowiki>  </nowiki>21.2
 
 
 
  |
 
<nowiki>  </nowiki>10.6
 
 
 
  |
 
<nowiki>  </nowiki>5.30 
 
 
|-
 
  |
 
<nowiki>  </nowiki>''' '''
 
 
 
  | 
 
 
 
  | 
 
 
 
  | 
 
 
 
  | 
 
 
 
  |   
 
 
|-
 
  |
 
<nowiki>  </nowiki>'''7'''
 
 
 
  |
 
<nowiki>  </nowiki>'''Trace metals'''
 
 
 
  |
 
<nowiki>  </nowiki>[M] g/mol
 
 
 
  |
 
<nowiki>  </nowiki>mg/L
 
 
 
  |
 
<nowiki>  </nowiki>mg/500 mL
 
 
 
  |
 
<nowiki>  </nowiki>mg/250 mL 
 
 
|-
 
  |
 
 
 
  |
 
<nowiki>  </nowiki>Na<sub>2</sub> · EDTA
 
 
 
  |
 
<nowiki>  </nowiki>372.24
 
 
 
  |
 
<nowiki>  </nowiki>4360
 
 
 
  |
 
<nowiki>  </nowiki>2 180
 
 
 
  |
 
<nowiki>  </nowiki>1 090.0 
 
 
|-
 
  | 
 
 
 
  |
 
<nowiki>  </nowiki>FeCl<sub>3</sub> · 6 H<sub>2</sub>O
 
 
 
  |
 
<nowiki>  </nowiki>270.32
 
 
 
  |
 
<nowiki>  </nowiki>315
 
 
 
  |
 
<nowiki>  </nowiki>1 575
 
 
 
  |
 
<nowiki>  </nowiki>787.5 
 
 
|-
 
  |
 
 
 
  |
 
<nowiki>  </nowiki>CuSO<sub>4</sub> · 5 H<sub>2</sub>O
 
 
 
  |
 
<nowiki>  </nowiki>249.69
 
 
 
  |
 
<nowiki>  </nowiki>10
 
 
 
  |
 
<nowiki>  </nowiki>5
 
 
 
  |
 
<nowiki>  </nowiki>2.5 
 
 
|-
 
  | 
 
 
 
  |
 
<nowiki>  </nowiki>ZnSO<sub>4</sub> · 7 H<sub>2</sub>O
 
 
 
  |
 
<nowiki>  </nowiki>287.54
 
 
 
  |
 
<nowiki>  </nowiki>22
 
 
 
  |
 
<nowiki>  </nowiki>11
 
 
 
  |
 
<nowiki>  </nowiki>5.5 
 
 
|-
 
  | 
 
 
 
  |
 
<nowiki>  </nowiki>CoCl<sub>2</sub> · 6 H<sub>2</sub>O
 
 
 
  |
 
<nowiki>  </nowiki>237.93
 
 
 
  |
 
<nowiki>  </nowiki>10
 
 
 
  |
 
<nowiki>  </nowiki>5
 
 
 
  |
 
<nowiki>  </nowiki>2.5 
 
 
|-
 
  | 
 
 
 
  |
 
<nowiki>  </nowiki>MnCl<sub>2</sub> · 4 H2O
 
 
 
  |
 
<nowiki>  </nowiki>197.90
 
 
 
  |
 
<nowiki>  </nowiki>180
 
 
 
  |
 
<nowiki>  </nowiki>90
 
 
 
  |
 
<nowiki>  </nowiki>45 
 
 
|-
 
  | 
 
 
 
  |
 
<nowiki>  </nowiki>NaMoO<sub>4</sub> · 2 H<sub>2</sub>O
 
 
 
  |
 
<nowiki>  </nowiki>241.95
 
 
 
  |
 
<nowiki>  </nowiki>6
 
 
 
  |
 
<nowiki>  </nowiki>3
 
 
 
  |
 
<nowiki>  </nowiki>1.5 
 
 
|-
 
  | 
 
 
 
  |
 
<nowiki>  </nowiki>H<sub>3</sub>BO<sub>3</sub>
 
 
 
  |
 
<nowiki>  </nowiki>61.83
 
 
 
  |
 
<nowiki>  </nowiki>1000
 
 
 
  |
 
<nowiki>  </nowiki>500
 
 
 
  |
 
<nowiki>  </nowiki>250.0 
 
 
|-
 
  | 
 
 
 
  | 
 
 
 
  |
 
 
 
  |
 
 
 
  |
 
 
 
  |   
 
 
|-
 
  |
 
<nowiki>  </nowiki>'''8'''
 
 
 
  |
 
<nowiki>  </nowiki>'''Vitamins'''
 
 
 
  |
 
<nowiki>  </nowiki>[M] g/mol
 
 
 
  |
 
<nowiki>  </nowiki>mg/L
 
 
 
  |
 
<nowiki>  </nowiki>mg/500 mL
 
 
 
  |
 
<nowiki>  </nowiki>mg/250 mL
 
 
|-
 
  | 
 
 
 
  |
 
<nowiki>  </nowiki>Thiamin · HCl
 
 
 
  |
 
<nowiki>  </nowiki>337.30
 
 
 
  |
 
<nowiki>  </nowiki>100.0
 
 
 
  |
 
<nowiki>  </nowiki>50.00
 
 
 
  |
 
<nowiki>  </nowiki>25.000 
 
 
|-
 
  | 
 
 
 
  |
 
<nowiki>  </nowiki>Biotin
 
 
 
  |
 
<nowiki>  </nowiki>244.31
 
 
 
  |
 
<nowiki>  </nowiki>0.5
 
 
 
  |
 
<nowiki>  </nowiki>0.25
 
 
 
  |
 
<nowiki>  </nowiki>0.125 
 
 
|-
 
  | 
 
 
 
  |
 
<nowiki>  </nowiki>B<sub>12</sub>
 
 
 
  |
 
<nowiki>  </nowiki>1355.38
 
 
 
  |
 
<nowiki>  </nowiki>0.5
 
 
 
  |
 
<nowiki>  </nowiki>0.25
 
 
 
  |
 
<nowiki>  </nowiki>0.125 
 
 
 
|}
 
|}
  
 
+
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 15:10, 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.