Aim: How can we analyze DNA?

1. Do Now: 1. What are the advantages of selective breeding? Give an example. Hw: Aim: How can we analyze DNA?

2. Pre-Lab Questions 1. What happens to DNA prior to electrophoresis? 2.How is DNA separated in gel electrophoresis? 3. How are the bands DNA arranged in the gel? Hw: Read pgs. 173-174 Answer Regents Practice B 12 on pg. 184, Part C 20 on pg. 185. Read pgs. 80 & 81 Define theory, fossil record, geologic time. Aim: How can we model the technique of gel electrophoresis?

3. Biotechnology Gel Electrophoresis

4. 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…

5. 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?

6. 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”

7. 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 Gel electrophoresis DNA  – + “swimming through Jello”

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

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

10. 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

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

12. 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

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

14. 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 +

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

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

17. 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”

18. 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”

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

20. I’m a-glow! Got any Questions?

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

22. 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

23. 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

24. 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

25. 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

26. 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