1. Contributing Authors: Pat Chrenka Alicia Harkins-Pritchett Melissa LaPlace Madeleine Martinez Emily Zablocki

2. Gel Electrophoresis Origins First created in 1970’s Created at Cold Spring Harbor Laboratory  Phil Sharp  Joe Sanbrook  Bill Sugden

3. 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

4. Required Equipment Agarose powder 1X TAE solution (buffer) 100 base pair ladder Ethidium bromide 1.5 mL test tube Micropipettes Micropipette tips Microwave Electrophoresis equipment Erlenmeyer flask Gloves *Simplified versions of this equipment can be used to make this more cost effective in your classrooms.

5. Step 1. Making the gel Mix 1.0 g agarose and 250 mL 1X TAE solution in Erlenmeyer flask. Stir solution

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

7. Add 4 microliters of Ethidium Bromide (wear protective gloves) to gel mixture.

8. Pour agarose solution into gel tray.

9. Place comb into liquid agarose.

10. Allow gel to solidify for approximately 15 minutes.

11. Step 2: Entering the DNA Fill electrophoresis chamber with 1X TAE solution. Make sure solution fills entire chamber and covers gel completely.

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

13. Add 25 microliters of DNA samples to other lanes.

14. Connect electrical current to gel electrodes. Run current for 20-30 minutes at 110 volts.

15. Step 3: Reading the Gel After the gel has been run, examine it under ultraviolet light to see the bands of DNA.

16. Lastly, listen as James explains how to determine whether or not Alu PV92 is part of your genetic make-up.

17. Gel Electrophoresis Model http://www.sumanasinc.com/webcontent/animations/content/gel electrophoresis.html

18. Forensic Science, Medical Science, Public Health, Food and Livestock Production  Exonerate persons wrongly accused of crimes  Identify crime and catastrophe victims  Establish paternity and other family relationships http://www.youtube.com/watch?v=SGtWssdauME  Identify potential suspects whose DNA may match evidence left at crime scenes  Identify 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 food  Match organ donors with recipients in transplant programs  Determine pedigree for seed or livestock breeds  Authenticate consumables such as caviar and wine How does this apply to society?

19. 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)

20. Applications in the Classroom Used during DNA units Used during forensic units Used during genetic and heredity units

21. Interactive websites for students http://learn.genetics.utah.edu/units/biotech/gel/http://learn.genetics.utah.edu/units/biotech/gel/ http://www.life.uiuc.edu/molbio/geldigest/electro.html#gel http://www.sumanasinc.com/webcontent/animations/c ontent/gelelectrophoresis.html