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Agarose Gel Electrophoresis with Dyes
Teacher Guide
Gel Electrophoresis: Diagnosis of Huntington’s Disorder
Some questions to get you thinking about today’s lab:
What is DNA and what does it do?
What are some characteristics or properties of DNA?
How can we take advantage of these properties to help us figure out genetic disorders?
Materials
- DNA samples Mom, Dad,(A) Patient A (Daughter), and (B) Patient B (Son)
- Molecular Weight Marker
- Electrophoresis units and power supplies
- agarose
- Tris-acetate/EDTA solution (TAE) or TBE
- micropipette/tips
1. Get your electrophoresis apparatus. Make sure the comb is in place and that there are stoppers at both ends of the gel space.
2. Pour hot agarose into the gel space until it reaches the top of the gel casting tray. Let the agarose harden, which should take about 10 minutes. Don't touch/move your gel until it's hard. Why not?
Draw a picture of your gel and label which samples are where before you add DNA to the gel.
3. Load your DNA SAMPLES into the wells near the BLACK ELECTRODE. Why near the black electrode? Be sure to keep track of which samples you loaded in which lanes.
4. Now pour TAE solution over your gel CAREFULLY so that is it completely covered plus a little more. What do you think the TAE solution is for?
5. Run that gel!! Plug the electrodes into your gel box (red to red, black to black), being careful not to bump your gel too much. Plug the power source into an outlet and set at 125 V. How can you tell your gel is running?
Analyzing Your DNA Data
Draw a picture of your gel.
Marker starts with 30nt then, 60, 90nt and120nt. Everything closer to the wells of the 120 nt marker is bigger than 120 nt.
Use the molecular weight marker information to determine the size of each person’s alleles. Based on the number of nucleotides in the region of DNA determine the number of trinucleotide repeats
Link to the Worksheet/Datasheet for the Electrophoresis Dyes lab
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BIOTECH Project Department of Molecular and Cellular Biology The University of Arizona August 3, 1998 Designed by: Ken Kubo, Ph.D. Last Modified March 1, 2002 Nadja Anderson, Ph.D. nadja@email.arizona.edu
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