Contributing Authors: Pat Chrenka Alicia Harkins-Pritchett Melissa LaPlace Madeleine Martinez Emily...
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Transcript of Contributing Authors: Pat Chrenka Alicia Harkins-Pritchett Melissa LaPlace Madeleine Martinez Emily...
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Contributing Authors:
Pat Chrenka
Alicia Harkins-Pritchett
Melissa LaPlace
Madeleine Martinez
Emily Zablocki
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Gel Electrophoresis Origins
First created in 1970’sCreated at Cold Spring Harbor LaboratoryPhil SharpJoe SanbrookBill Sugden
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How does it work?
Uses electricity to separate DNA fragments by size
Uses an electric current to separate different size molecules in a porous sponge-like matrix
Larger molecules move more slowly than smaller molecules
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Required Equipment
Agarose powder1X TAE solution (buffer)100 base pair ladderEthidium bromide 1.5 mL test tubeMicropipettesMicropipette tipsMicrowaveElectrophoresis equipmentErlenmeyer flaskGloves
*Simplified versions of this equipment can be used to make this more cost effective in your classrooms.
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Step 1. Making the gel
Mix 1.0 g agarose and 250 mL 1X TAE solution in Erlenmeyer flask.Stir solution
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Heat solution in microwave. Use short 20 to 30 second heatings. Swirl after each heating. Repeat
until solution is clear.
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Add 4 microliters of Ethidium
Bromide (wear protective gloves)
to gel mixture.
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Pour agarose solution into gel tray.
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Place comb into liquid agarose.
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Allow gel to solidify for
approximately 15 minutes.
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Step 2: Entering the DNA
Fill electrophoresis chamber with
1X TAE solution. Make sure solution
fills entire chamber and
covers gel completely.
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Load 5 microliters of DNA ladder to first lane of gel.
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Add 25 microliters of DNA samples to other lanes.
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Connect electrical
current to gel electrodes. Run current
for 20-30 minutes at 110
volts.
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Step 3: Reading the Gel
After the gel has been run, examine it under ultraviolet light to see the bands of DNA.
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Lastly, listen as James explains how to determine whether or not Alu PV92
is part of your genetic make-up.
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Gel Electrophoresis Model
http://www.sumanasinc.com/webcontent/animations/content/gelelectrophoresis.html
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Forensic Science, Medical Science, Public Health, Food and Livestock Production
Exonerate persons wrongly accused of crimesIdentify crime and catastrophe victimsEstablish paternity and other family relationships
http://www.youtube.com/watch?v=SGtWssdauME Identify potential suspects whose DNA may match evidence left at
crime scenesIdentify endangered and protected species as an aid to wildlife officials
(could be used for prosecuting poachers)Detect bacteria and other organisms that may pollute air, water, soil,
and foodMatch organ donors with recipients in transplant programsDetermine pedigree for seed or livestock breedsAuthenticate consumables such as caviar and wine
How does this apply to society?
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What did we learn from this experience?
I learned how to read the base pair bands of the DNA sample on the gel. (ML)
This has been a great review from undergrad biology and how I can use this in my classroom. (AHP)
I learned that technology I previously thought was restricted to scientists, has practical application in the middle school classroom. (MM)
I learned the variety of uses of gel electrophoresis which impact daily life. I can now take these examples to my classroom and bring real life and science together for my students. (EZ)
We have the ability to take science from the laboratory and present in the classroom to facilitate learning experiences for students. (PC)
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Applications in the Classroom
Used during DNA unitsUsed during forensic unitsUsed during genetic and heredity
units
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Interactive websites for students
http://learn.genetics.utah.edu/units/biotech/gel/
http://www.life.uiuc.edu/molbio/geldigest/electro.html#gel
http://www.sumanasinc.com/webcontent/animations/content/gelelectrophoresis.html