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
8:30
8:45
9:30
9:45
Location
Planetarium
CRW230
&231
Planetarium
CRW230
&231
10:30 Planetarium
CRW230
10:45 &231
11:30 Quarterdeck
12:30 Planetarium
1:00 Planetarium
CRW230
1:15 &231
2:00 Planetarium
2:15 Planetarium
2:30
Activity
Introduction
1) DNA Extraction & Genes in a
Bottle
PCR Overview
15 min
45 min
15 min
2) Food color pipetting & Virtual PCR
Gel and Capillary ElectrophoresisTour
45 min
3) Gel Electrophoresis
Lunch
4) Restriction Mapping
Transformation Overview
45 min
45 min
30 min
15 min
5) pGLO Transformation
The future of Molecular Biology?
molecules in fossils?
Wrap-up other talks?
Depart LSSU
45 min
15 min
15 min
15 min
*Log on to the following website and and answer these questions:
AP Biology Restriction Enzymes
http://www.phschool.com/science/biology_place/biocoach/red/intro.html
1.
2.
3.
4.
5.
Explain what a restriction enzyme is and what it does.
Describe what a restriction fragment is and how the are formed.
Give 2 reasons why restriction fragments are important in gene technology.
Explain what a restriction site is.
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 
http://www.phschool.com/science/biology_place/labbench/lab6/concepts2.html
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.