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Go to the following website http://learn.genetics.utah.edu/ Go to the tab for "Virtual Labs". You will be doing the DNA extraction and the Gel Electrophoresis. Complete this worksheet as you walk through the steps of the labs. DNA Extraction 1. List three reasons for extracting DNA from a human subject: 1) ________________________________________________________ 2) _________________________________________________________ 3) __________________________________________________________ 2. Where is DNA found in the cell? _____________________ 3. From where do we obtain the cells of our test subject? ___________________________ 4. List the four steps needed to extract DNA: 1) ________________________________________________________ 2) _________________________________________________________ 3) __________________________________________________________ 4) __________________________________________________________ 5. The lysis solution contains detergent, what will this do to the cell? ________________________________ It also contains proteinase K, what does this do to the cell? ________________________________________ 6. What does the salt do to the cellular mixture? _________________________________ 7. Why do you need to place a second tube into the centrifuge? ______________________________ After the tube is removed from the centrifuge, what equipment is used to remove the top liquid from the tube? _____________________________________________ What is in this liquid? ____________________________________ 8. Isopropyl alcohol causes DNA to do what? _________________________________ Gel Electrophoresis 1. Gel electrophoresis is used to sort DNA strands according to their __________________ Where do you place the DNA samples on the gel? _______________________________ What makes the DNA move? _____________________ Short strands move [ faster / slower ] than longer strands. 2. Place the steps in the correct order (number them) ______ Load DNA sample into the gel ______ Stain the Gel and analyze results ______ Make the gel ______ Hook up the electrical current ______ Set up gel apparatus 3. In the "Gel Electrophoresis Laboratory", follow the steps to make your own gel, answer the questions as you go. a) What is agarose made from? ___________________________________ b) Melted agarose is poured into a ___________________________________ c) What is the purpose of the comb? ___________________________________________ d) The black end generates a ______________ charge, the red end a _______________ charge. e) The bubbles in the electrophoresis indicate what? _________________________________ f) Staining the DNA will make it show up under a ____________________________________ light. g) What are your estimates for the number of base pairs in the three bands? ___________________ DNA Fingerprinting Site: PBS / NOVA - go to http://www.pbs.org/wgbh/nova/ , click on Nova Teachers, then go to Life Science and find the activity on creating a DNA fingerprint. Direct link: http://www.pbs.org/wgbh/nova/teachers/body/createdna-fingerprint.html. You can also search for this using google by searching for "create a dna fingerprint" Objective: Students will learn the steps of DNA fingerprinting by creating a fingerprint in a virtual lab. They will use this fingerprint to solve a virtual crime. The virtual lab is interactive and goes through the step-by-step process of DNA fingerprinting 1. What crime was committed? 2. Who are the suspects? Creating a DNA Fingerprint 3. What is Step 1? 4. What does a restriction enzyme do to DNA? 5. What is Step 2? 6. Describe agarose: 7. What is Step 3? 8. What is Step 4? 9. What is electrophoresis? 10. Which fragments of DNA will move the farthest? 11. What is Step 5? 12. What is the purpose of the nylon membrane? 13. What is Step 6? 14. How are the probes labeled? 15. What is Step 7? 16. Analyze the DNA. Which one of the suspects is the culprit? Site 2: Go to DNA.gov 17. Browse the website until you get a good idea what the purpose of the site is. In your opinion, what is DNA.gov being used for? 18. DNA can be used to solve crimes, but the government also uses it for what other purposes? 19. Name a course that is offered at this site. 20. Look under statistics and find how many offender profiles are stored in your state's database. What is a database? Web Lesson: Cloning Learn.Genetics at http://learn.genetics.utah.edu/ -- > Under the "Cell Biology" Menu, Click on "Cloning" Browse the articles at the site to find the answers to the following questions. What is Cloning? 1. Define Cloning: _________________________________________________ 2. What is the difference between natural twinning and artificial twinning? ______________________________________ 3. What is SCNT? ____________________________________________ 4. To make Dolly the clone, they first isolated a cell from where? __________________________________ 5. They transferred the nucleus of this cell to where? ____________________ Click and Clone 6. List all the materials needed to clone a mouse. 7. Place the following steps in the correct order. ________ Stimulate cell division ________ Deliver baby ________ Remove and discard the nucleus from the egg cell ________ Isolate donor cells from egg donor and germ cell donor ________ Transfer the somatic cell nucleus into the egg cell ________ Implant embryo into a surrogate mother 8. Explain how the nucleus is removed from the donor egg: 9. What color with the cloned mouse be? _____________ What is the name of this mouse? _____________________ The History of Cloning 10. What was the first organism ever cloned using artificial embryo twinning? ___________________________________________ 11. How did Hans Sermann separate cells from a salamander embryo? _______________________________ 12. What happened to the tadpoles that were cloned from more advanced embryos? ______________________________ 16. Where did John Gurdon obtain cells for his cloned frogs? _________________________________ 17. What was the first mammal embryo cloned? ________________________________ 18. What were the names of the two cloned calves?____________________________________ 19. Cultured sheeps cells were used to create two lambs named _______________________ and ____________________ 20. What as the first mammal created using somatic cell nuclear transfer? _________________________ Where did the DNA come from for this clone? ______________________ 21. Out of 29 primate clones attempted, how many were born? _______ 22. How was the clone "Polly" different from the first sheep clone named "Dolly?" __________________________ 23. What was the first extinct animal cloned? ____________________________________ Why Clone? 24. List four reasons given on the page for justifyig cloning: _________________________________________________ _________________________________________________ __________________________________________________ __________________________________________________ 25. What is a stem cell ? ___________________________________________ 26. Why did CC look different from Rainbow even though they were both clones? _________________________________ 27. What two things would you need to clone a dinosaur? _______________________________________________ 28. Why would a farmer want to clone livestock? ___________________________________________ 29. What are two reasons a person might want to clone a human? _____________________________________ Is it Cloning or Not? 30. For each of the following scenarios, indicate YES (it is cloning) or NO (it is not cloning) ___________Sperm taken from a mole goat is combined with a female's egg in a petri dish. The resulting embryo is implanted into the female's uterus to develop ___________A sheep embryo, composed of 16 cells, is removed from the mother's uterus and separated into indivudal cells. Each cell is allowed to multiply, creating 16 separate embryos, which are then implanted in different female sheep to develop to maturity. ___________A cow with many desirable traits is stimulated with hormones to produce a number of egg cells. Each of these eggs is fertilized and implanted into a surrogate mother. ___________ In vitro fertilization ___________ Cell nuclei from an extinct wolly mammoth are placed into enucleated cow cells. Cloning Myths 31. Why is it impossible to make instant clones? _____________________________________ 32. Why would Frankie #2 be different from Frankie #1? ____________________________________________ 33. Give an example of a natural clone: _____________________________________________ 34. Humans have been cloning _______________________ for thousands of years. 35. Why does cloning have such a high failure rate? _______________________________________________ Review the following site to aid in understanding more on biotechnology. https://docs.google.com/presentation/d/1zZAW6j0qp3DpQOaSGY0dIT9fKlOpt5iS3gKiTZ6TetE/present?sl ide=id.i180