Sigma xi presentation slides

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1. Portable InfraredSystem for AmplifyingDNA andConducting FieldNucleic Acid TestingPRESENTER: DILLON MIR 2. Hypothesis The use of infrared light to heat PCR tubes in order to create a simplified and cost effective method of DNA amplification. 3. Hypothesis questions Can we create a model of PCR tube heating using infrared lamps? Can we create a heating device that can be bought for a low cost or easily made? Can we make this device easy to use? 4. Research methods First tests were run with water in order to create the heating model without using an excess of the somewhat expensive DNA helicase. Tests were run starting at room temperature (~24C). Lowered or elevated temperatures causes a difference time to heat the tube. PCR tube heated to about 65 degrees Celsius. The heating and cooling models of different temperatures were created in order to find a time and voltage the would not damage the DNA sample. Allowed to cool to room temperature before another experiment was run. 5. The first prototypeInfraredBreadboardlampsAluminumPCR tubeblock 6. Heating Equations and derivation 7. Heating Equations and derivation 8. Heating Equations and derivation 9. Heating Equations and derivation 10. Heating Equations and derivation 11. Cooling Equations and derivation 12. Cooling Equations and derivation 13. Cooling Equations and derivation 14. Cooling Equations and derivation 15. Cooling Equations and derivation 16. For elevated temperatures 17. For elevated temperatures 18. For elevated temperatures 19. Prototype 2 The next prototype we created was a Sun of Vergina design. Holds up to 6 PCR tubes for multiple tests at once. Made of clay with silver paint for reflection. 20. Prototype 2 InfraredBreadboard lamps2ndprototypePCR tubes 21. Future prototype Created in Solidworks. Once designed it can be sent to a machine shop to be created. Easy to produce more with the same specifications. 22. DNA amplification procedures DNA amplification process: 1. Combine necessary components together in a PCR tube. 2. Heat PCR tube for 30 minutes. 3. Remove liquid from PCR tube and place in glass slide. 4. turn out light (except blue side LED). 5. Observe drop (green color in drop is amplified DNA). 23. DNA amplification MaterialsQuantity (L) Water14 10X Annealing Buffer 2.5 MgSO41 NaCl 2 IsoAmp dNTP Solution 2 DNA template 2.5 Forward Primer 1 Reverse Primer 1 IsoAmp Enzyme Mix2 24. DNA amplification Blue light makes for easier analysis using image software. Green color inside drop is the amplified (copied) DNA. Higher concentrations of amplified DNA will produce a more visible shade of green. 25. Results The data values are the temperature that was measured with respect to time at different voltages. The prediction values are based on the heating model that was created from the collected data. 26. Conclusions Using infrared lamps to heat PCR tubes was successful. Heating and cooling models of a PCR tube with reasonable accuracy were created. A multi-tube DNA amplification platform was successfully designed and tested. 27. Acknowledgements Dr. Antonio Garcia Instructor/Advisor Professor Karmella Haynes Provided DNA helicase samples Angel Lastra Equations and derivations