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What Colors Your World?

What Colors Your World?. Biotechnology Activities with Food Dyes. Instructors - Bio-Rad Curriculum and Training Specialists. Sherri Andrews, Ph.D., Eastern US sherri_andrews@bio-rad.com Damon Tighe, Western US damon_tighe@bio-rad.com Leigh Brown, M.A., Central US

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What Colors Your World?

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  1. What Colors Your World? Biotechnology Activities with Food Dyes

  2. Instructors - Bio-Rad Curriculum and Training Specialists Sherri Andrews, Ph.D., Eastern US sherri_andrews@bio-rad.com Damon Tighe, Western US damon_tighe@bio-rad.com Leigh Brown, M.A., Central US leigh_brown@bio-rad.com

  3. Separation technology: How do you separate two or more compounds from each other?

  4. Combining Science, Engineering and Math to Develop a Separation Technology Engineering – Design parameter based; no single “correct answer” Science – Hypothesis driven; try to find specific cause/effect Best separation material? Measurements Analysis Understanding the chemistry of mobile phase- Salt? Polar? Non-polar? Inexpensive separates well

  5. Chromatography • Used to separate biomolecules based in their physical characteristics may include: • size • charge • hydrophobicity • interaction with other molecules

  6. The System • All forms of chromatography have a • Stationary phase (remains stationary and is a solid or a liquid supported by a solid) • Mobile phase (a liquid or a gas that travels through the stationary phase and carries the parts of the mixture)

  7. Types of Chromatography • Paper • Paper serves as the stationary phase. • The paper strip is placed in a solvent (mobile) phase which carries the mixture through the paper. • Parts of the mixture will travel at different rates separating the parts • Column • The stationary phase is packed into a column • The column is equilibrated (saturated with solvent) • Sample is added to the column • Buffers are added to separate the mixture • Samples are collected in intervals called fractions

  8. Be an Engineer – Design your own! • Select materials that you will use to separate your Kool Aid into the food dyes that make it the color it appears • You will do this by • Paper chromatography • Column chromatography

  9. What are some of the design factors we want to think about?

  10. Dye separation from Kool Aid Using Paper Chromatography • Choose your paper type (stationary phase) • Cut it into .75 x 4 inch strips • Place 50ul of Kool-Aid at one end using a DPTP

  11. Place 1 ml of your mobile phase into a medicine cup • Place the strip Kool-Aid side down into the mobile phase

  12. Building and Using A Column to Separate Your Kool-Aid Choose your Column Type (HIC, Empty Column, Syringe) If empty column or syringe add stationary phase by stuffing it with a cotton ball

  13. Building and Using A Column to Separate Your Kool-Aid • Choose your mobile phase (water, alcohol, or 1xPBS) • Place column in collection tube • Add 2 ml of mobile phase to column • Let it flow until it no longer drips

  14. Building and Using A Column to Separate Your Kool-Aid • Move column to next collection tube • Using a DPTP add 1ml of Kool-Aid to the top of the column • Let it flow until it no longer drips (this is fraction 1) • Move to the next column • Continue steps 5 and 6 until the eluate (fluid coming off the column) is clear

  15. Compare Results Which paper, mobile phase worked best for paper chromatography? Which column, mobile phase worked best for column chromatography?

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