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Biotechnology Gel Electrophoresis

Biotechnology Gel Electrophoresis. Many uses of restriction enzymes…. Now that we can cut DNA with restriction enzymes… we can cut up DNA from different people… or different organisms… and compare it why? forensics medical diagnostics paternity evolutionary relationships and more….

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Biotechnology Gel Electrophoresis

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  1. Biotechnology Gel Electrophoresis

  2. Many uses of restriction enzymes… • Now that we can cut DNA with restriction enzymes… • we can cut up DNA from different people… or different organisms… and compare it • why? • forensics • medical diagnostics • paternity • evolutionary relationships • and more…

  3. Comparing cut up DNA • How do we compare DNA fragments? • separate fragments by size • How do we separate DNA fragments? • run it through a gelatin • gel electrophoresis • How does a gel work?

  4. Gel electrophoresis • A method of separating DNA in a gelatin-like material using an electrical field • DNA is negatively charged • when it’s in an electrical field it moves toward the positive side DNA        – + “swimming through Jello”

  5. Gel electrophoresis • DNA moves in an electrical field… • so how does that help you compare DNA fragments? • size of DNA fragment affects how far it travels • small pieces travel farther • large pieces travel slower & lag behind DNA        – + “swimming through Jello”

  6. Gel Electrophoresis DNA &restriction enzyme - longer fragments wells power source gel shorter fragments completed gel +

  7. fragments of DNAseparate out based on size Running a gel Stain DNA • ethidium bromide binds to DNA • fluoresces under UV light cut DNA with restriction enzymes 1 2 3

  8. DNA fingerprint • Why is each person’s DNA pattern different? • sections of “junk” DNA • doesn’t code for proteins • made up of repeated patterns • CAT, GCC, and others • each person may have different number of repeats • many sites on our 23 chromosomes with different repeat patterns GCTTGTAACGGCCTCATCATCATTCGCCGGCCTACGCTT CGAACATTGCCGGAGTAGTAGTAAGCGGCCGGATGCGAA GCTTGTAACGGCATCATCATCATCATCATCCGGCCTACGCTT CGAACATTGCCGTAGTAGTAGTAGTAGTAGGCCGGATGCGAA

  9. Allele 1 cut sites repeats cut sites GCTTGTAACGGCCTCATCATCATTCGCCGGCCTACGCTT CGAACATTGCCGGAGTAGTAGTAAGCGGCCGGATGCGAA GCTTGTAACG GCCTCATCATCATCGCCG GCCTACGCTT CGAACATTGCCG GAGTAGTAGTAGCGGCCG GATGCGAA DNA patterns for DNA fingerprints Cut the DNA 1 2 3 – + DNA allele 1

  10. Differences between people Person 1 cut sites cut sites GCTTGTAACGGCCTCATCATCATTCGCCGGCCTACGCTT CGAACATTGCCGGAGTAGTAGTAAGCGGCCGGATGCGAA Person 2: more repeats GCTTGTAACGGCCTCATCATCATCATCATCATCCGGCCTACGCTT CGAACATTGCCGGAGTAGTAGTAGTAGTAGTAGGCCGGATGCGAA 1 2 3 DNA fingerprint – + DNA person 1 person 2

  11. 1 2 3 4 5 1 2 3 4 5 Uses: Evolutionary relationships • Comparing DNA samples from different organisms to measure evolutionary relationships turtle snake rat squirrel fruitfly – DNA  +

  12. Uses: Medical diagnostic • Comparing normal allele to disease allele chromosomewith normal allele 1 chromosome with disease-causing allele 2 allele 2 allele 1 – DNA  Example: test for Huntington’s disease +

  13. Uses: Forensics • Comparing DNA sample from crime scene with suspects & victim suspects crime scene sample S1 S2 S3 V – DNA  +

  14. DNA fingerprints • Comparing blood samples on defendant’s clothing to determine if it belongs to victim • DNA fingerprinting

  15. RFLP / electrophoresis use in forensics • 1st case successfully using DNA evidence • 1987 rape case convicting Tommie Lee Andrews “standard” semen sample from rapist blood sample from suspect “standard” How can you compare DNA fromblood & from semen?RBC? “standard” semen sample from rapist blood sample from suspect “standard”

  16. Electrophoresis use in forensics • Evidence from murder trial • Do you think suspect is guilty? blood sample 1 from crime scene blood sample 2 from crime scene blood sample 3 from crime scene “standard” blood sample from suspect OJ Simpson blood sample from victim 1 N Brown blood sample from victim 2 R Goldman “standard”

  17. Mom F1 F2 child Uses: Paternity • Who’s the father? – DNA  +

  18. I’m a-glow! Got any Questions?

  19. Biodiversity Lab Which plant is most closely related to Botanus curus?

  20. Botanus curus Sequences CAC GTG GAC TGA GGA CTC CTC DNA GUG CAC CUG ACU CCU GAG GAG RNA Val His Leu Thr Pro Glu Glu AA

  21. Species X Sequences CAC GTG GAC TGA GGA CTC CTC BC CAC GTG GAC AGA GGA CAC CTC DNA GUG CAC CUG UCU CCU GUG GAG RNA Val His Leu Ser Pro Val Glu AA Val His Leu Thr Pro Glu Glu BC

  22. Species Y Sequences CAC GTG GAC TGA GGA CTC CTC BC CAC GTG GAC AGA GGA CAC CTC DNA GUG CAC CUG UCU CCU GUG GAG RNA Val His Leu Ser Pro Val Glu AA Val His Leu Thr Pro Glu Glu BC

  23. Species Z Sequences CAC GTG GAC TGA GGA CTC CTC BC CAC GTA GAC TGA GGA CTT CTC DNA GUG CAU CUG ACU CCU GAA GAG RNA Val His Leu Thr Pro Glu Glu AA Val His Leu Thr Pro Glu Glu BC

  24. Amino Acid Sequences Val His Leu Thr Pro Glu Glu BC Val His Leu Ser Pro Val Glu X Val His Leu Ser Pro Val Glu Y Val His Leu Thr Pro Glu Glu Z

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