Team:WashU/Protocols/ZeaxanthinExtraction

From 2012.igem.org

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1. Take cells and spin them down at 10,000 g's for 15 minutes.
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1. Take cells and spin them down at 10,000 g's for 15 minutes. <br>
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2. Discard supernatant.
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2. Discard supernatant.<br>
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3. Resuspend pellet in ethanol using approximately 5% of the original culture volume. Ensure that pellet is fully resuspended. Vortex and mash to mix.
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3. Resuspend pellet in ethanol using approximately 5% of the original culture volume. Ensure that pellet is fully resuspended. Vortex and mash to mix.<br>
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4. Add the same volume of hexane to your solution.
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4. Add the same volume of hexane to your solution.<br>
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5. Vortex and shake vigoriously for one minute.
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5. Vortex and shake vigoriously for one minute.<br>
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6. Centrifuge at 10,000 g's for 5 minutes.
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6. Centrifuge at 10,000 g's for 5 minutes.<br>
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7. Remove organic layer (which will contain the yellow color) and place it in a new test tube with one boiling chip and submerge in 70 degree water bath until the organic solvent is driven off. NOTE: Do this in a fume hood, as hexane vapors will be emitted from the vaporizing solution.
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7. Remove organic layer (which will contain the yellow color) and place it in a new test tube with one boiling chip and submerge in 70 degree water bath until the organic solvent is driven off. NOTE: Do this in a fume hood, as hexane vapors will be emitted from the vaporizing solution.<br>
CAUTION: The hexane will superheat if not careful. Thus, handle with caution (use hot hands if necessary, as well).
CAUTION: The hexane will superheat if not careful. Thus, handle with caution (use hot hands if necessary, as well).
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[https://2012.igem.org/Team:WashU/Protocols Back to Protocols]

Latest revision as of 22:05, 9 August 2012


1. Take cells and spin them down at 10,000 g's for 15 minutes.
2. Discard supernatant.
3. Resuspend pellet in ethanol using approximately 5% of the original culture volume. Ensure that pellet is fully resuspended. Vortex and mash to mix.
4. Add the same volume of hexane to your solution.
5. Vortex and shake vigoriously for one minute.
6. Centrifuge at 10,000 g's for 5 minutes.
7. Remove organic layer (which will contain the yellow color) and place it in a new test tube with one boiling chip and submerge in 70 degree water bath until the organic solvent is driven off. NOTE: Do this in a fume hood, as hexane vapors will be emitted from the vaporizing solution.

CAUTION: The hexane will superheat if not careful. Thus, handle with caution (use hot hands if necessary, as well).

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