Instructor Terry Wiseth

1. IF YOU CAN SEE THIS MESSAGE YOU ARE NOT IN “SLIDE SHOW” MODE. PERFOMING THE LAB IN THIS MODE WILL NOT ALLOW FOR THE ANIMATIONS AND INTERACTIVITY OF THE EXERCISE TO WORK PROPERLY. TO CHANGE TO “SLIDE SHOW” MODE YOU CAN CLICK ON “VIEW” AT THE TOP OF THE PAGE AND SELECT “SLIDE SHOW” FROM THE PULL DOWN MENU. YOU CAN ALSO JUST HIT THE “F5” KEY. Instructor Terry Wiseth Start Lab

2. Ether You can use the links below to view the data sheets, punnett squares and questions you will need to hand in for this lab. You can also begin the lab by using the links on the right. Eye Color Data Sheet Go to Introduction f1 Eye Color Punnett Square Wing Formation Data Sheet f2 Eye Color Punnett Square Skip Introduction and go directly to collecting data Eye Color, Wing Formation Data Sheet f1 Wing Formation Punnett Square Genetics Questions 1-5 f2 Wing Formation Punnett Square PUNNET SQUARE TUTORIAL (if needed) f1 Eye Color, Wing Formation Punnett Square f2 Eye Color, Wing Formation Punnett Square Next Slide Previous Slide End Lab

3. Ether Click on the blackboard to zoom in to a larger view of the blackboard Drosophila melanogaster is a fruit fly, a little insect about 3mm long, of the kind that accumulates around spoiled fruit. It is also one of the most valuable of organisms in biological research, particularly in genetics and developmental biology. Drosophila has been used as a model organism for research for almost a century, and today, several thousand scientists are working on many different aspects of the fruit fly. Brush Ether Fly Cultures Time Machine Next Slide Previous Slide End Lab Skip Intro

4. Ether Part of the reason people work on it is historical. So much is already known about it that it is easy to handle and well understood. Part of it is practical: it's a small animal, with a short life cycle of just two weeks, and is cheap and easy to keep in large numbers. Mutant flies, with defects in any of several thousand genes are available, and the entire genome has recently been sequenced. Next Slide Previous Slide End Lab Skip Intro

5. Ether The drosophila egg is about half a millimeter long. It takes about one day after fertilization for the embryo to develop and hatch into a worm-like larva. The larva eats and grows continuously, molting numerous times after hatching. After about 6 days as a larva, it molts one more time to form an immobile pupa. Over the next 4 days, the body is completely remodeled to give the adult winged form, which then hatches from the pupal case and is fertile within about 12 hours. Next Slide Previous Slide End Lab Skip Intro

6. Ether It is essential that you learn to distinguish the males from the females. With practice this becomes relatively easy. In older flies the posterior part of the abdomen is quite dark in males and considerably lighter in females. The tip of the abdomen is more rounded in males than in females. In general the male is smaller than the female. Female Male Females are slightly larger than males Posterior abdomen is darker than females Next Slide Previous Slide End Lab Skip Intro

7. Ether MAKING CROSSES:The strains of flies we are using are all true breeding so are assumed to be homozygous. You will be concerned only with the traits listed on the vial for your flies. “+” is used to represent the wild type flies. r + Next Slide Previous Slide End Lab Skip Intro

8. Ether The wild type (“+”) flies will exhibit normal phenotypes and genotypes. Normal phenotypes include full length wings and red eyes. The mutant strains of these characteristics that we will be working with include curly wings and brown eyes. Our first experiment will include a monohybrid cross between normal “+” red-eyed flies and mutant brown-eyed flies. Our objectivewill be to determine phenotypicand genotypic ratios as well as dominant and recessive genes. Next Slide Previous Slide End Lab Skip Intro

9. Ether You will need to use a Punnet square to assist in developing possible genotypes. In most cases the mutant strain of a characteristic is recessive and the normal or wild-type is dominant. The crosses that we will be performing and carrying to the f2 generation are as follows: 1) Red Eye X Brown Eye 2) Full Wing X Curly Wing 3) Red Eye, Full Wing X Brown Eye, Curly Wing If you are unfamiliar with using aPunnet Square you can use thislink to perform aPUNNET SQUARE TUTORIAL Next Slide Previous Slide End Lab Skip Intro

10. Ether For your crosses and Punnet squares, we will use the following abbreviations: R = Red Eye r = Brown Eye F = Full Wing f = Curly Wing If you are unfamiliar with using aPunnet Square you can use thislink to perform aPUNNET SQUARE TUTORIAL Next Slide Previous Slide End Lab Skip Intro

11. Ether You can use the links below to view the data sheets, punnett squares and questions you will need to hand in for this lab. Eye Color Data Sheet, f1 Punnet Square, f2 Punnet Square Wing Formation Data Sheet, f1 Punnet Square, f2 Punnet Square Eye Color, Wing Formation Data Sheet, f1 Punnet Square, f2 Punnet Square Virtual Fly Lab Questions 1-5 PUNNET SQUARE TUTORIAL (if needed) Next Slide Previous Slide End Lab

12. Ether In order to begin the lab we will need to obtain breeding males and females. You will need to capture 3 virgin females and 3 males from different culture bottles. Click on the fly cultures to bring two culture bottles to the table. Then Click on NEXT SLIDE Brush Ether Fly Cultures r + Time Machine ON Next Slide Previous Slide End Lab Introduction

13. Ether One of the culture bottles is labeled as “+” which indicates the wild type or normal homozygous red-eyed genotype and phenotypes. The other culture bottle is labeled as “r” for a mutant homozygous brown-eyed genotype and phenotype. Click on NEXT SLIDE Brush Ether Fly Cultures r + Time Machine ON Next Slide Previous Slide End Lab

14. Ether Ether To acquire the adult flies we will need to anesthetize them and combine the chosen flies in an new culture bottle. Click on the microscope and the ether anesthesia to bring them to the table. Then Click on NEXT SLIDE Brush Ether Fly Cultures r + Time Machine ON Next Slide Previous Slide End Lab

15. Click on the handle of the Ether wand to anesthetize the flies in both culture bottles. Be very careful to leave the Ether wand in the culture bottle for only a few seconds so that you do not kill or sterilize the flies. It will take about 30 seconds for the flies to go to sleep. Click on NEXT SLIDE Brush Ether Fly Cultures Ether r + Time Machine ON Next Slide Previous Slide End Lab

16. + Ether + When the flies are no longer moving you can click on the “+” culture bottle to empty the flies onto the microscope plate. Click on the turned culture bottle to remove the sponge stopper and to tap the flies onto the microscope stage. Brush Ether Fly Cultures r + Time Machine ON Next Slide Previous Slide End Lab

17. + + Ether Next Click on the NEXT SLIDE button When the flies are no longer moving you can click on the “+” culture bottle to empty the flies onto the microscope plate. Click on the turned culture bottle to remove the sponge stopper and to tap the flies onto the microscope stage. Brush Ether Fly Cultures r Time Machine ON Next Slide Previous Slide End Lab

18. Ether P1 + Click on the used “+” type culture tube on the desk in order to dispose of it. Next click on the fly culture tubes on the shelf to obtain a fresh culture tube for your new cross. Label this tube as P1. Then click on NEXT SLIDE. Brush Ether Fly Cultures r Time Machine ON Next Slide Previous Slide End Lab

19. Ether P1 Click on the microscope to get a magnified image of your “+” type flies. Click on the sponge plug on the P1 culture bottle to remove it. You will need to Click on three females to add to the fresh breeding culture for the monohybrid cross. Click on NEXT SLIDE when you are done This is not a FEMALE This is not a FEMALE This is not a FEMALE This is not a FEMALE This is not a FEMALE Brush Ether Fly Cultures r Time Machine Next Slide Previous Slide End Lab

20. Ether r P1 Click on the NEXT SLIDE button. Now we will harvest 3 males from the brown-eyed culture (r). Click on the culture tube labeled “r” to empty the anesthetized flies on the microscope stage. Next Click on the turned culture bottle to remove the sponge stopper and to tap the flies onto the stage. Brush Ether Fly Cultures r + Time Machine ON Next Slide Previous Slide End Lab

21. r Ether Click on the microscope to get a magnified image of your “r” type flies. Click on the sponge plug on the P1 culture bottle to remove it. You will need to Click on three males to add to the fresh breeding culture for the monohybrid cross. Click on NEXT SLIDE when you are done This is not a MALE This is not a MALE This is not a MALE This is not a MALE Brush Ether Fly Cultures + P1 Time Machine Next Slide Previous Slide End Lab

22. Ether Click on the sponge plug to replace on the P1 culture bottle. Your flies should wake up and begin to move around. We will need to wait for 10 days in order for the parent flies to mate and the offspring to emerge from their pupae. Click on the NEXT SLIDE button. Brush Ether Fly Cultures + P1 Time Machine ON Next Slide Previous Slide End Lab

23. Ether Fortunately we are living at a time when we have the use of a Time Machine. We only need to insert our culture tube into the Time Machine and we can fast forward to 10 days in the future. This is much better than waiting for 10 days to find our results. Click on NEXT SLIDE. Brush Ether Fly Cultures + P1 Time Machine ON Next Slide Previous Slide End Lab

24. Ether + P1 Click on the P1 culture tube to insert it into the Time Machine. When the machine stops we will be 10 days in the future and we will be able view the offspring of our parental flies. After the tube is placed Click the yellow ON switch for the Time Machine Brush Ether Fly Cultures + P1 Time Machine ON Next Slide Previous Slide End Lab

25. Ether + P1 When the Time Machine has stopped, the culture tube will be returned to the table (10 days later). Click on the Ether Wand to anesthetize the offspring from your cross. Click on the P1 culture tube to empty the flies onto the microscope stage. Now Click on NEXT SLIDE. Brush Ether Fly Cultures + P1 Time Machine ON Next Slide Previous Slide End Lab

26. + P1 When the Time Machine has stopped, the culture tube will be returned to the table (10 days later). Click on the Ether Wand to anesthetize the offspring from your cross. Click on the P1 culture tube to empty the flies onto the microscope stage. Now Click on NEXT SLIDE. Brush Ether Fly Cultures Ether + P1 Time Machine ON Next Slide Previous Slide End Lab

27. + P1 When the Time Machine has stopped, the culture tube will be returned to the table (10 days later). Click on the Ether Wand to anesthetize the offspring from your cross. Click on the P1 culture tube to empty the flies onto the microscope stage. Now Click on NEXT SLIDE. Brush Ether Fly Cultures + P1 Ether Time Machine ON Next Slide Previous Slide End Lab

28. P1 Click on the Microscope to observe a zoomed in view of the f1 offspring. In this view count the number of red-eyed flies and brown-eyed flies and record on your data sheet. Click on NEXT SLIDE when you are finished counting. In this view count the number of red-eyed flies and brown-eyed flies and record on your data sheet as the f1 generation. Click on NEXT SLIDE when you are finished counting. Brush Ether Fly Cultures Ether Time Machine ON BrownEyes RedEyes Next Slide Previous Slide End Lab

29. Click on the Microscope to observe a zoomed in view of the P1 flies. In this view count the number of red-eyed flies and brown-eyed flies and record on your data sheet. Click on NEXT SLIDE when you are finished counting. Click on a male fly and a female fly to insert the flies into a new culture tube this time labeled as f1 Brush Ether Fly Cultures Ether Time Machine ON BrownEyes RedEyes Next Slide Previous Slide End Lab

30. We now have a red-eyed f1 culture which will be allowed to produce an f2 generation which we can analyze. Click on the f1 culture tube to insert the culture into the Time Machine. Brush Ether Fly Cultures Ether + f1 Time Machine ON Next Slide Previous Slide End Lab

31. Ether + f1 After the tube is placed Click the yellow ON switch for the Time Machine Brush Ether Fly Cultures + P1 Time Machine ON Next Slide Previous Slide End Lab

32. Ether + f1 When the Time Machine has stopped, the culture tube will be returned to the table (10 days later). Click on the Ether Wand to anesthetize the offspring from your cross. Click on the f1 culture tube to empty the flies onto the microscope stage. Now Click on NEXT SLIDE. Brush Ether Fly Cultures + f1 Time Machine ON Next Slide Previous Slide End Lab

33. + f1 When the Time Machine has stopped, the culture tube will be returned to the table (10 days later). Click on the Ether Wand to anesthetize the offspring from your cross. Click on the f1 culture tube to empty the flies onto the microscope stage. Now Click on NEXT SLIDE. Brush Ether Fly Cultures Ether + P1 Time Machine ON Next Slide Previous Slide End Lab

34. + f1 When the Time Machine has stopped, the culture tube will be returned to the table (10 days later). Click on the Ether Wand to anesthetize the offspring from your cross. Click on the f1 culture tube to empty the flies onto the microscope stage. Now Click on NEXT SLIDE. Brush Ether Fly Cultures + f1 Ether Time Machine ON Next Slide Previous Slide End Lab

35. f1 Click on the Microscope to observe a zoomed in view of the f2 offspring. In this view count the number of red-eyed flies and brown-eyed flies and record on your data sheet. Click on NEXT SLIDE when you are finished counting. In this view count the number of red-eyed flies and brown-eyed flies and record on your data sheet as the f2 generation. Click on NEXT SLIDE when you are finished counting. Brush Ether Fly Cultures Ether Time Machine ON BrownEyes RedEyes Next Slide Previous Slide End Lab

36. Curly Wings Next Slide Previous Slide End Lab

37. Ether Next we will investigate the inheritance of normal Full Wings vs Curly Wings. Again you will need to capture females and males from different culture bottles. Click on the fly cultures to bring two culture bottles to the table. Then Click on NEXT SLIDE Brush Ether Fly Cultures f + Time Machine ON Next Slide Previous Slide End Lab

38. Ether One of the culture bottles is labeled as “+” which indicates the wild type or normal homozygous full winged genotype and phenotypes. The other culture bottle is labeled as “f” for a mutant homozygous curly winged genotype and phenotype. Click on NEXT SLIDE Brush Ether Fly Cultures f + Time Machine ON Next Slide Previous Slide End Lab

39. Ether Ether To acquire the adult flies we will need to anesthetize them and combine the chosen flies in an new culture bottle. Click on the microscope and the ether anesthesia to bring them to the table. Then Click on NEXT SLIDE Brush Ether Fly Cultures f + Time Machine ON Next Slide Previous Slide End Lab

40. Click on the handle of the Ether wand to anesthetize the flies in both culture bottles. Be very careful to leave the Ether wand in the culture bottle for only a few seconds so that you do not kill or sterilize the flies. It will take about 30 seconds for the flies to go to sleep. Click on NEXT SLIDE Brush Ether Fly Cultures Ether f + Time Machine ON Next Slide Previous Slide End Lab

41. + Ether + When the flies are no longer moving you can click on the “+” culture bottle to empty the flies onto the microscope plate. Click on the turned culture bottle to remove the sponge stopper and to tap the flies onto the microscope stage. Brush Ether Fly Cultures f + Time Machine ON Next Slide Previous Slide End Lab

42. + + Ether Next Click on the NEXT SLIDE button When the flies are no longer moving you can click on the “+” culture bottle to empty the flies onto the microscope plate. Click on the turned culture bottle to remove the sponge stopper and to tap the flies onto the microscope stage. Brush Ether Fly Cultures f Time Machine ON Next Slide Previous Slide End Lab

43. Ether P1 + Click on the used “+” type culture tube on the desk in order to dispose of it. Next click on the fly culture tubes on the shelf to obtain a fresh culture tube for your new cross. Label this tube as P1. Then click on NEXT SLIDE. Brush Ether Fly Cultures f Time Machine ON Next Slide Previous Slide End Lab

44. Ether P1 Click on the microscope to get a magnified image of your “+” type flies. Click on the sponge plug on the P1 culture bottle to remove it. You will need to Click on three females to add to the fresh breeding culture for the monohybrid cross. Click on NEXT SLIDE when you are done This is not a FEMALE This is not a FEMALE This is not a FEMALE This is not a FEMALE This is not a FEMALE Brush Ether Fly Cultures f Time Machine Next Slide Previous Slide End Lab

45. Ether f P1 Now we will harvest 3 males from the curly winged culture (f). Click on the culture tube labeled “f” to empty the anesthetized flies on the microscope stage. Next Click on the turned culture bottle to remove the sponge stopper and to tap the flies onto the stage. Click on the NEXT SLIDE button. Brush Ether Fly Cultures f + Time Machine ON Next Slide Previous Slide End Lab

46. f Ether Click on the microscope to get a magnified image of your “f” type flies. Click on the sponge plug on the P1 culture bottle to remove it. You will need to Click on three males to add to the fresh breeding culture for the monohybrid cross. Click on NEXT SLIDE when you are done This is not a MALE This is not a MALE This is not a MALE Brush Ether Fly Cultures + P1 Time Machine Next Slide Previous Slide End Lab

47. Ether Click on the sponge plug to replace on the P1 culture bottle. Your flies should wake up and begin to move around. We will need to wait for 10 days in order for the parent flies to mate and the offspring to emerge from their pupae. Click on the NEXT SLIDE button. Brush Ether Fly Cultures + P1 Time Machine ON Next Slide Previous Slide End Lab

48. Ether + P1 Click on the P1 culture tube to insert it into the Time Machine. When the machine stops we will be 10 days in the future and we will be able view the offspring of our parental flies. After the tube is placed Click the yellow ON switch for the Time Machine Brush Ether Fly Cultures + P1 Time Machine ON Next Slide Previous Slide End Lab

49. Ether + P1 When the Time Machine has stopped, the culture tube will be returned to the table (10 days later). Click on the Ether Wand to anesthetize the offspring from your cross. Click on the P1 culture tube to empty the flies onto the microscope stage. Now Click on NEXT SLIDE. Brush Ether Fly Cultures + P1 Time Machine ON Next Slide Previous Slide End Lab

50. + P1 When the Time Machine has stopped, the culture tube will be returned to the table (10 days later). Click on the Ether Wand to anesthetize the offspring from your cross. Click on the P1 culture tube to empty the flies onto the microscope stage. Now Click on NEXT SLIDE. Brush Ether Fly Cultures Ether + P1 Time Machine ON Next Slide Previous Slide End Lab