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Gel Electrophoresis

Gel Electrophoresis. Ryan Yang and Violet Kamber 6 th Period ITC. AGENDA. Group Development Timeline Problem Statement Background/Justification Customer Scope Deliverables Brainstorming Research and Generate Ideas. Identify Criteria and Constraints Explore Possibilities

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Gel Electrophoresis

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  1. Gel Electrophoresis Ryan Yang and Violet Kamber 6th Period ITC

  2. AGENDA • Group Development • Timeline • Problem Statement • Background/Justification • Customer • Scope • Deliverables • Brainstorming • Research and Generate Ideas • Identify Criteria and Constraints • Explore Possibilities • Develop a Design Proposal • Select an approach • Prototype • Test and Monitor • Refine • Lesson Learned • Summary

  3. GROUP DEVELOPEMENT • Role and Responsibilities: • Team Leader: Ryan Yang • Time Keeper: Violet Kamber • Scribe: Ryan Yang • Team Constitution:

  4. TIMELINE • Completion dates confirmed: • 10 days 1/27-2/8 • 7 days 2/6-2/22 • 8 days 2/22-3/5

  5. PROBLEM STATEMENT • To catch a criminal whose DNA was left at the scene, a Gel Electrophoresis Chamber will be created to test the DNA and figure out whether any of the 3 suspects were involved. This is so that we can catch the culprit and recover a valuable piece of artwork.

  6. BACKGROUND • Gel Electrophoresis: • Uses restriction enzymes • Helps catch suspect • Separates DNA

  7. CUSTOMERS • Mr. Pritchard • Guiding through the process • Mr. Church • Guiding through DNA testing • Museum • Where artwork was stolen

  8. SCOPE • Build a Gel Electrophoresis Tester • Gel carrier, chamber, lid, and comb • Removable power source • Submit technical report • Submit PowerPoint

  9. SCOPE • Utilize Sources • Internet, library, teachers • Technical – Mr. Pritchard • Inventor – Mr. Hund • Calculations/ Math – Ms. Hernacki

  10. SCOPE Materials needed: • Plexiglass • Adhesive/sealant • Banana plugs • Wires • Agarose gel • Buffer • Drill press • Bandsaw • Screws • Plastic (comb)

  11. SCOPE Constraints: - Gel must be 3/8” high - Teeth 1/8”+ from wall of Carrier - Carrier must hold all 50 mL

  12. DELIVERABLES • Technical report • Purpose, planning, organization • PowerPoint • Prototype • Fits criteria and constraints

  13. BRAINSTORMING Questions: • Dimensions for the carrier? • Composition of the buffer? • Analyzing DNA? • How much voltage? • Composition of gel?

  14. BRAINSTORMING • Use PLTW 12-Step • Use 50 ml of Agarose • Agarose 3/8 inches tall • DNA separated by size • Restriction enzymes to cut DNA pieces • Different enzymes have different bases • A power supply is connected to box • Direct current is applied to the samples. • The higher voltage, faster molecules travel

  15. RESEARCH AND GENERATE IDEAS Possible dimensions for the carrier? HxWxD (cm) • 0.9525 x 10 x 5.25 • 0.9525 x 7.5 x 7 • 0.9525 x 8 x 6.5625 • 0.9525 x 8.75 x 6 • 0.9525 x 10.5 x 5

  16. RESEARCH AND GENERATE IDEAS Composition of buffer(TBE)? • 218g Tris base • 110g Boric acid • 9.3g EDTA • 1.9L of distilled water What is the composition of the gel? • 0.5g of agarose • 50ml of TBE • 1µL of ethidium bromide

  17. RESEARCH AND GENERATE IDEAS Ideal voltage for the gel? • Distance between the electrodes 5V per each cm Analyze the separated DNA? • Estimate molecular weights by comparing bands to DNA markers

  18. RESEARCH AND GENERATE IDEAS Key Ideas: • Gel is heated and cooled • 30 minutes to an hour to set • Wells filled with DNA and loading buffer • The final lane should be filled with marker • Current checked to ensure flowing

  19. EXPLORE POSSIBILITIES Solution 1

  20. EXPLORE POSSIBILITIES • Box dimensions: 5.442 x 4.442 x 1.5 • Lid Dimensions: Base : 4.884 x 5.442 x .221 • 2 panels: .5 x 5.442 x .221 • Carrier Dimensions: Base: 2.801 x 3.44 x .221 • Walls: .221 x 3.44 x .75 • Comb Dimensions: Straight Side: 2.801 x .125 x .125 • Pegs: .375 x .125 x .125

  21. EXPLORE POSSIBILITIES Solution 2

  22. EXPLORE POSSIBILITIES • Box Dimensions: 3.992 x 5.5 x 1.821 • Lid Dimensions: Base 5.942 x 3.922 x .221 • 2 Panels: 3.922 x .221 x .279 • Carrier Dimensions: Base : 2.509 x 3.94 x .221 • Walls: 3.94 x .221 x .625 • Comb Dimensions: Straight Side: 3.44 x .125 x .125 • Pegs: .125 x .125 x .313

  23. EXPLORE POSSIBILITIES Solution 3

  24. EXPLORE POSSIBILITIES • Box Dimensions: 4.442 x 4.442 x 1.5 • Lid Dimensions: Base: 4.884 x 4.442 x .221 • 2 Panels: .221 x 4.442 x .5 • Carrier Dimensions: Base: 2.953 x 3.198 x .221 • Walls: .221 x 2.953 x .625 • Comb Dimensions: Straight Side: 3.198 x .125 x .125 • Pegs: .276 x .125 x .25

  25. SELECT AN APPROACH Solution • Yellow and Orange have tied • Hybridization of Solution 1 and Solution 3 • Solution 3 has higher scores • Solution 1 highest Dimensional Fitness and best Voltage to Distance Ratio

  26. DESIGN PROPOSAL • Build Process • Worked

  27. REFINE • Counter-boredhole lid to shut off power • Lid should be wider • Guarantee safety. • Comb adjusted 3/16th length to accommodate DNA and buffer

  28. LESSONS LEARNED • Planning ahead is a key • Effectively distribute tasks • Murphy’s Law will come into play • Effectively get building done • Separate sections of the pieces • Sealant was applied in high amount • Not too much excess sealant • Work together

  29. SUMMARY • Good learning experience • Overcome obstacles • Success despite some delays • Get ahead in case of Murphy’s Law • Extra time was spent • Building took approximately 5-7 days • Design planned and adjusted for strength

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