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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

Gel Electrophoresis

Ryan Yang and Violet Kamber

6th Period ITC


Agenda
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


GROUP DEVELOPEMENT

  • Role and Responsibilities:

    • Team Leader: Ryan Yang

    • Time Keeper: Violet Kamber

    • Scribe: Ryan Yang

  • Team Constitution:


Timeline
TIMELINE

  • Completion dates confirmed:

  • 10 days 1/27-2/8

  • 7 days 2/6-2/22

  • 8 days 2/22-3/5


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.


Background
BACKGROUND

  • Gel Electrophoresis:

    • Uses restriction enzymes

    • Helps catch suspect

    • Separates DNA


CUSTOMERS

  • Mr. Pritchard

    • Guiding through the process

  • Mr. Church

    • Guiding through DNA testing

  • Museum

    • Where artwork was stolen


Scope
SCOPE

  • Build a Gel Electrophoresis Tester

    • Gel carrier, chamber, lid, and comb

    • Removable power source

    • Submit technical report

    • Submit PowerPoint


Scope1
SCOPE

  • Utilize Sources

    • Internet, library, teachers

    • Technical – Mr. Pritchard

    • Inventor – Mr. Hund

    • Calculations/ Math – Ms. Hernacki


Scope2
SCOPE

Materials needed:

  • Plexiglass

  • Adhesive/sealant

  • Banana plugs

  • Wires

  • Agarose gel

  • Buffer

  • Drill press

  • Bandsaw

  • Screws

  • Plastic (comb)


Scope3
SCOPE

Constraints:

- Gel must be 3/8” high

- Teeth 1/8”+ from wall of Carrier

- Carrier must hold all 50 mL


Deliverables
DELIVERABLES

  • Technical report

    • Purpose, planning, organization

  • PowerPoint

  • Prototype

    • Fits criteria and constraints


Brainstorming
BRAINSTORMING

Questions:

  • Dimensions for the carrier?

  • Composition of the buffer?

  • Analyzing DNA?

  • How much voltage?

  • Composition of gel?


Brainstorming1
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


Research and generate ideas
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


Research and generate ideas1
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


Research and generate ideas2
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


Research and generate ideas3
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



Explore possibilities1
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



Explore possibilities3
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



Explore possibilities5
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


Select an approach
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


Design proposal
DESIGN PROPOSAL

  • Build Process

    • Worked


Refine
REFINE

  • Counter-boredhole lid to shut off power

  • Lid should be wider

  • Guarantee safety.

  • Comb adjusted 3/16th length to accommodate

    DNA and buffer


Lessons learned
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


Summary
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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