Gel Electrophoresis Experiment Fall 2007

1. Gel Electrophoresis Experiment Fall 2007 A Timed Presentation - do not click the mouse. Approximate Run Time - 4 minutes 5 seconds

2. Comb produces 32 wells Gel Apparatus Gel is 22.5 x 22.5 cm in size

3. Restriction Enzyme Digest Oven Temperature accuracy is good to + or - 1 degree celsius

4. Two students mixing the DNA and restriction Enzymes

5. Rabieh is setting up for the gel electrophoresis experiment

6. Simon Loading Lane 1

7. Ian Loading Lane 2

8. Ryan Loading Lane 3

9. Katen Loading Lane 4

10. Joshua Loading Lane 5

11. Andrew Loading Lane 6

12. Patty Loading Lane 7

13. Sara Loading Lane 8

14. Jessica Loading Lane 9

15. Chantal Loading Lane 10

16. Steve Loading Lane 11

17. Alborz Loading Lane 12

18. Jessica Loading Lane 13

19. Alicia Loading Lane 14

20. Emily Loading Lane 15

21. Megan Loading Lane 16

22. Lindsey Loading Lane 17

23. Stephanie Loading Lane 18

24. Danielle Loading Lane 19

25. Erika Loading Lane 20

26. Hailey Loading Lane 21

27. Ashmita Loading Lane 22

28. Rachel Loading Lane 23

29. Linda Loading Lane 24

30. Sameera Loading Lane 25

31. Rabieh Loading Lane 26 - OR NOT?

32. Erika Loading Lane 27

33. Sumaiya Loading Lane 28

34. Loading the DNA was a success!

35. Checking the gel before turning on the power

36. The class is a busy place during the time it takes to load the gel

38. Protocol Uncut: DNA 18.75 μl 10 x Buffer 15 μl Water 116.25 μl 6 x Loading Buffer 30 μl 30 μl/well – approximately 1.25 μg DNA/well Each sample of DNA was incubated for 1 hour at 37°C. Each sample and the ladder was then loaded into each well as outlined in the following slides. Single cut: DNA 37.5 μl 10 x Buffer 22.5 μl BamHI 15 μl Water 150 μl 6 x Loading Buffer 45 μl 30 μl/well – approximately 1.67 μg DNA/well BamHI 900 bp pBabe/cpp32 6000 bp EcoRI Bouble cut: DNA 37.5 μl 10 x Buffer 22.5 μl BamHI 11.25 μl EcoRI 11.25 μ l Water 142.5 μl 6 x Loading Buffer 45 μl 30 μl/well – approximately 1.67 μg DNA/well 5100 bp pBabe plasmid donated by Dr. Tang

39. Due to some technical difficulty not yet figured out, the gel did not run as it should have The following results are obtained from a previous lab

40. DNA Ladder used in the gel electrophoresis experiments

41. Ladder Rows: 1 8 18 28 Background light

42. 2 To 7 Uncut Plasmid Rows Background light

43. Single Digest Rows 9 to 17 HindIII Background light

44. Double Digest Rows 19 to 27 Background light

45. 400 bp

46. 850 bp

47. 1650 bp

48. 2000 bp

49. 5000 bp

50. 6000 bp