Contributing Authors:

Pat Chrenka

Alicia Harkins-Pritchett

Melissa LaPlace

Madeleine Martinez

Emily Zablocki

Gel Electrophoresis Origins

First created in 1970’sCreated at Cold Spring Harbor LaboratoryPhil SharpJoe SanbrookBill Sugden

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

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.

Step 1. Making the gel

Mix 1.0 g agarose and 250 mL 1X TAE solution in Erlenmeyer flask.Stir solution

Heat solution in microwave. Use short 20 to 30 second heatings. Swirl after each heating. Repeat

until solution is clear.

Add 4 microliters of Ethidium

Bromide (wear protective gloves)

to gel mixture.

Pour agarose solution into gel tray.

Place comb into liquid agarose.

Allow gel to solidify for

approximately 15 minutes.

Step 2: Entering the DNA

Fill electrophoresis chamber with

1X TAE solution. Make sure solution

fills entire chamber and

covers gel completely.

Load 5 microliters of DNA ladder to first lane of gel.

Add 25 microliters of DNA samples to other lanes.

Connect electrical

current to gel electrodes. Run current

for 20-30 minutes at 110

volts.

Step 3: Reading the Gel

After the gel has been run, examine it under ultraviolet light to see the bands of DNA.

Lastly, listen as James explains how to determine whether or not Alu PV92

is part of your genetic make-up.

Gel Electrophoresis Model

http://www.sumanasinc.com/webcontent/animations/content/gelelectrophoresis.html

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?

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)

Applications in the Classroom

Used during DNA unitsUsed during forensic unitsUsed during genetic and heredity

units

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