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<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>. |
+ | == 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" |
+ | ! | ||
+ | ! | ||
+ | ! | ||
+ | ! | ||
+ | |- | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |- | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |- | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |} | ||
+ | {| class="MsoTableGrid" | ||
+ | |||
+ | | colspan="2" | | ||
+ | <nowiki> </nowiki>'''Buffer''' | ||
+ | |||
+ | | | ||
+ | <nowiki> </nowiki>[M] g/mol | ||
+ | |||
+ | | | ||
+ | <nowiki> </nowiki>mg/5 L | ||
+ | |||
+ | | | ||
+ | <nowiki> </nowiki>mg/L | ||
+ | |||
+ | | | ||
+ | |||
+ | |- | ||
+ | | | ||
+ | |||
+ | | | ||
+ | <nowiki> </nowiki>TES | ||
+ | |||
+ | | | ||
+ | <nowiki> </nowiki>229.25 | ||
+ | |||
+ | | | ||
+ | <nowiki> </nowiki>575 | ||
+ | |||
+ | | | ||
+ | <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 | ||
+ | |||
+ | |} | ||
== References == | == References == |
Revision as of 17:11, 18 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[1], and the most-commonly used recipe includes modifications as reported by Makulla[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 |
[M] g/mol |
mg/5 L |
mg/L |
||
TES |
229.25 |
575 |
115 |
||
Stock solutions |
[M] g/mol |
g/L |
g/500 mL |
g/250 mL | |
1 |
CaCl2 · 2 H2O |
147.02 |
36.8 |
18.4 |
9.20 |
2 |
MgSO4 · 7 H2O |
246.48 |
37.0 |
18.5 |
9.25 |
3 |
NaHCO3 |
84.01 |
12.6 |
6.3 |
3.15 |
4 |
K2HPO4 |
174.18 |
8.7 |
4.4 |
2.18 |
5 |
NaNO3 |
85.01 |
85.0 |
42.5 |
21.25 |
6 |
NaSiO3 · 5 H2O |
212.16 |
21.2 |
10.6 |
5.30 |
|
|||||
7 |
Trace metals |
[M] g/mol |
mg/L |
mg/500 mL |
mg/250 mL |
Na2 · EDTA |
372.24 |
4360 |
2 180 |
1 090.0 | |
FeCl3 · 6 H2O |
270.32 |
315 |
1 575 |
787.5 | |
CuSO4 · 5 H2O |
249.69 |
10 |
5 |
2.5 | |
ZnSO4 · 7 H2O |
287.54 |
22 |
11 |
5.5 | |
CoCl2 · 6 H2O |
237.93 |
10 |
5 |
2.5 | |
MnCl2 · 4 H2O |
197.90 |
180 |
90 |
45 | |
NaMoO4 · 2 H2O |
241.95 |
6 |
3 |
1.5 | |
H3BO3 |
61.83 |
1000 |
500 |
250.0 | |
8 |
Vitamins |
[M] g/mol |
mg/L |
mg/500 mL |
mg/250 mL |
Thiamin · HCl |
337.30 |
100.0 |
50.00 |
25.000 | |
Biotin |
244.31 |
0.5 |
0.25 |
0.125 | |
B12 |
1355.38 |
0.5 |
0.25 |
0.125 |