AP Biology Molecular Biology Lab Worksheet—Electrophoresis



AP Biology Molecular Biology Lab Worksheet—Electrophoresis/Restriction Enzymes 4-16-13

*Here is our schedule for Friday’s Molecular Biology Lab. We are building the base information for these procedures this week. We did the PCR SG yesterday. Today we will do restriction enzymes and gel electrophoresis. Remember we will all meet in the Crawford Hall Lobby at 8:30…..DO NOT BE LATE!! As always…..I love you guys especially when you are on time. (

|Time |Location |Activity | |

|8:30 |Planetarium |Introduction |15 min |

|8:45 |CRW230 &231 |1) DNA Extraction & Genes in a Bottle |45 min |

|9:30 |Planetarium |PCR Overview |15 min |

|9:45 |CRW230 &231 |2) Food color pipetting & Virtual PCR |45 min |

|10:30 |Planetarium |Gel and Capillary Electrophoresis-Tour |15 min |

|10:45 |CRW230 &231 |3) Gel Electrophoresis |45 min |

|11:30 |Quarterdeck |Lunch |45 min |

|12:30 |Planetarium |4) Restriction Mapping |30 min |

|1:00 |Planetarium |Transformation Overview |15 min |

|1:15 |CRW230 &231 |5) pGLO Transformation |45 min |

|2:00 |Planetarium |The future of Molecular Biology? |15 min |

| | |molecules in fossils? | |

|2:15 |Planetarium |Wrap-up other talks? |15 min |

|2:30 | |Depart LSSU | |

*Log on to the following website and and answer these questions:

AP Biology Restriction Enzymes



1. Explain what a restriction enzyme is and what it does.

2. Describe what a restriction fragment is and how the are formed.

3. Give 2 reasons why restriction fragments are important in gene technology.

4. Explain what a restriction site is.

5. Draw a picture of the 3 ways restrictions enzymes cleave fragments of DNA in your notes.

6. Explain what an Eppendorf tube is.

7. Give the 4 substances needed to do a restriction enzyme reaction.

8. At what temperature must this reaction take place and for how long?

9. What is the purpose of gel electrophoresis?

10. Briefly explain how the gel is made for this process.

11. What is the purpose of the buffer placed on top of the gel?

12. What 2 substances are added to the DNA in order to be able to track it?

13. Which way does the DNA move once the power is turned on ?

14. Explain why the shortest fragments of DNA move the farthest in the gel.

**Go through concept 4 until you understand it…….it is critical to the lab we will do Friday. We will go over it in class but you need to grasp the basics before we do (



15. Explain the function of restriction endonucleases. Give another name for them.

16. Explain what a recognition sequence is.

17. How do DNA engineers us restriction enzymes?

18. What is the main purpose of gel electrophoresis?

19. What will be done with the fragments of DNA separated in this experiment?

20. Explain what it means for restriction enzymes to be highly specific.

21. Explain the term palindromic as it applies to restriction enzymes.

22. Explain the function of the EcoRI enzyme.

**Hit the cut buttons on all the strands of DNA

23. Explain what a sticky end is and how they are formed.

24. Explain what a blunt end is and how they are formed.

25. Explain what is in the 20 microliter solution we will begin this phase of the experiment with. What temperature?

*Gel electrophoresis

26. How are molecules separated in gel electrophoresis?

27. Explain what affects the rate of movement of these molecules.

28. Explain what affects the speed with which they move.

29. Explain why current is applied and what affect is has on the molecules.

30. Which fragments of DNA move fastest? Slowest? Why?

31. Explain the 2 reasons for the tracking dye in this experiment.

32. How is the length of these DNA fragments measured?

33. Give the three types of DNA we will use and what organism they will come from.

34. Explain what Hind III is and what it does.

35. What will happen once these fragments are separated?

36. Explain why the gel must be stained after its run. How will it be stained?

37. Explain what a marker is and it’s purpose.

38. What is the marker in this experiment?

39. Explain what a standard curve is and how it is produced.

40. Explain what is meant by interpolation.

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