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Published byAbel Robinson Modified over 9 years ago
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DNA Fingerprinting Comparing DNA samples using gel electrophoresis
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How is DNA fingerprinting technology useful? Food identification Mystery meat Endangered species Forensics Crime scenes Overturning erroneous convictions Human remains
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Paternity/maternity Child support Immigration Health and medicine Genetic disorders Bone marrow transplants Identifying pathogens
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How can we examine DNA to look for similarities and differences? Electrophoresis “carry with electricity”
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DNA samples from crime scene and suspects
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Bacteria provide tools to cut DNA “DNA scissors” produced by bacteria Adaptation to protect bacteria against viruses
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Restriction Enzymes More than 3000 known restriction enzymes, or “endonucleases” Each cuts at a specific DNA sequence called a restriction site. Restriction sites are palindromes
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How do restriction enzymes work?
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What happens when we cut all DNA samples with the same restriction enzyme? 1 -------------GAATTC----------------------------- 2 ---GAATTC--------------------GAATTC------- 3 ---GAATTC----GAATTC----GAATTC------- 4 -----------------------------GAATTC------------- 1 -------------GA ATTC----------------------------- 2 ---GA ATTC--------------------GA ATTC------- 3 ---GA ATTC----GA ATTC----GA ATTC------- 4 ---------------------------------GA ATTC---------
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Electrophoresis animation Click here
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How will the fragments appear in a gel? 1 -------------GA ATTC----------------------------- 2 ---GA ATTC--------------------GA ATTC------- 3 ---GA ATTC----GA ATTC----GA ATTC------- 4 ---------------------------------GA ATTC---------
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Electrophoresis Procedure Cut DNA samples with restriction enzymes Add loading dye to samples Load samples in gel and apply charge DNA separates by size (small=fast) Stain DNA in the gel Bands appear where same-sized fragments are located, forming a pattern Compare patterns to find matches
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Pipette technique Find well opening with tip Depress plunger s..l..o..w..l..y… Watch for air bubbles New tip for each sample!
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The disposable graduated pipette
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