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The Phot oblea ch i ng Eff ect by Stephen Payne ?

Learn how to address photobleaching in DNA fluorescence with an LED resistor change from 4.3 Ohm to 20 Ohm, reducing the bleaching effect and enhancing signal quality in double-stranded DNA imaging. This adjustment optimizes the system for improved performance. For more details and acknowledgements, refer to the provided resources.

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The Phot oblea ch i ng Eff ect by Stephen Payne ?

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  1. ThePhotobleachingEffect by Stephen Payne ?

  2. Binds to double-stranded DNA and fluoresces Does not bind to single-stranded DNA SYBR Green

  3. The Problem LED Double-Stranded DNA Bleached SYBR Green Light Fluorescent SYBR Green

  4. Suspicion of The Problem

  5. Bleaching Function -.00078t Flu = e

  6. Bleaching Function -.00068t Flu = e

  7. Correction by Bleaching Factor

  8. Insignificant Bleaching Effect?

  9. The Solution • Change R1 in LED Driver from a 4.3 Ohm resistor to a 20 Ohm resistor

  10. Reduced Bleaching Effect

  11. The Theory 20 Ohms Optimal Signal Signal 4.3 Ohms R Resistance (Ohms) optimal

  12. Conclusion • Photobleaching significantly affected our system with the 4.3 Ohm resistor • This effect was reduced via increasing the resistance in the LED driver • Further optimization of the system is possible

  13. Acknowledgements Code • Danielle Carpenter, Co-author • Steve Wasserman, Debugging Images • http://distancelearning.ksi.edu/demo/bio378/DNA_files/image023.jpg • http://en.wikivisual.com/images/0/04/NA-comparedto-DNA_thymineAndUracilCorrected.png • http://www.elixa.com/light/arrays2.jpg • http://openwetware.org/wiki/20.309:Measuring_DNA_Melting_Curves

  14. Questions?

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