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DNA Fingerprinting

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DNA Fingerprinting
This lab must be typed. Title DNA Fingerprinting Purpose Why are we doing this lab Background 1. What are restriction enzymes 2. When added to a DNA sample, what do restriction enzymes do 3. What do you call the specific sequence of bases the enzyme is searching for 4. What is a restriction digestion 5. What is the purpose of the water bath 6. The electrophoresis apparatus creates an electrical field with positive and negative poles at the ends of the gel. DNA molecules are negatively charged. To which electrode pole of the electrophoresis field would you expect DNA to migrate Explain. 7. What color represents the negative pole 8. After DNA samples are loaded into the sample wells, they are forced to move through the gel matrix. What size fragments (large vs. small) would you expect to move toward the opposite end of the gel most quickly Explain. 9. Which fragments (large vs. small) are expected to travel the shortest distance from the well Explain. Materials List everything you used in the lab. Procedure List steps according to the handout. Day 1 and Day 2 Day 1 Observations 1. Describe the DNA samples. 2. Describe the restriction endonuclease mix. 3. Can you see any evidence to indicate that your samples of DNA were fragmented or altered in any way by the addition of EcoRl/Pstl Explain. 4. In the absence of any visible evidence of change, is it still possible that the DNA samples were fragmented Explain your reasoning. 5. After the restriction digest incubation period, are there any visible clues that the restriction enzymes may have in some way changed the DNA in any of the tubes Explain your reasoning. Day 2 Post Lab Observation of Results Sketch your result to the left. Dry your gel as instructed. Quantitative Analysis of DNA Fragment Sizes If you were on trial or were trying to identify an endangered species, would you want to rely on a technicians eyeball estimate of a match, or would you want some more accurate measurement In order to make the

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