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Download Gel Electrophoresis DNA Fingerprinting
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Transcript
Gel Electrophoresis DNA Fingerprinting DNA Analysis • How are DNA molecules analyzed. • Restriction enzyme digestion of DNA molecules. • Gel electrophoresis to separate DNA fragments of different sizes. Restriction Enzymes Where did they come from? • Bacteria! • Natural defense against viral infections • Cut up DNA at various bases sequences • May leave a “stickey end” • May leave “blunt end” Restriction Digestion of DNA Blunt End Sticky End Restriction Map Comparison DNA Analysis • How are DNA molecules analyzed. • Restriction Nuclease digestion of DNA molecules. • Gel electrophoresis to separate DNA fragments of different sizes. DNA Gel Electrophoresis QuickTime™ and a TIFF (LZW) decompressor are needed to see this picture. Today’s laboratory: DNA fingerprinting in a hypothetical forensics team investigation A Criminal Case • In this hypothetical case, DNA was extracted from samples obtained from the five possible suspects, and the crime scene sample • You will cleave the DNA with a restriction enzyme and simulated a “mock” DNA fingerprint analysis using Southern Blotting Step One: Cut up your DNA! Crime Scene Suspect Suspect One Two Step Two: Load Up Your Gel Look at those RFLP’s! Crime Scene Suspect One Suspect Two Step Three: It’s Blotting Time Step Four: Hybridize using DNA probe Step Five: Take a picture! Typical Southern Blot Gel Electrophoresis vs. Southern Blots Electrophoresis • Shows you ALL DNA fragments that result from restriction enzyme digest • All possible RFLP’s are shown Southern Blots • Additional step after electrophoresis • Looks from specific short DNA sequence inside of RFLP’s • Uses radioactive material • Much more specific than gels.
