Lesson plan #2

A.J. Pettway/Debbie Bell

7th Grade Life Science
7-18-03

Forensic Files: A DNA Dilemma

I. Anticipatory Set
At the beginning of the class period, students will find case files on their desks that they are to read.
The instructor will then explain that the file is from the LAPD and that the investigators there need the students help to
solve a murder case.

II. Background
Students must have a firm grasp of DNA and heredity
Basic knowledge of enzymes and what they do is a plus.
Graphing Skills
Students will benefit by having an understanding of the uniqueness of an individuals DNA sequence

III. Abstract
The purpose of this lab activity is to demonstrate (through simulation) how DNA fingerprinting (or DNA profiling) might be
used to solve a crime. In this activity, students perform restriction digests on DNA samples from four individuals, and then
search for similarities between the individuals by running the restriction fragments on an electrophoresis gel. Because not
two people ( except identical twins) have exactly the same DNA, a person's DNA fingerprint is unique and can be used for
purposes of identification. This activity does not do a true DNA fingerprint. It simulates two of the three steps of DNA
fingerprinting: restriction of DNA sample and separation by electrophoresis.

IV. Lesson Plan

A. Goal:
The lesson is designed to expose students to a relatively new DNA technology and how it is used in a practical setting to
identify individuals through DNA fingerprinting.

B. Content Objectives:
Through this activity students will learn: some properties of DNA, how Restriction Digestion uses enzymes to cut DNA, the
process of separating DNA based on its size (gel electrophoresis), basic measurement technique, how to generate and use
a standard curve, and the scientific method of investigation.

C. Materials:

4 ul. EcoRI enzyme

2 ul. BamHI enzyme

2 ul. HindIII enzyme

12 ul. lambda DNA (0.5 mg/ml)

20 ul. 2x multicore restriction buffer

2 ul. loading dye

1Kb DNA ladder

1% agarose gel(1g. agarose/ 100 ml. TBE or TAE buffermicrowave and pour)

1 liter 1X TBE or TAE Buffer

Carolina Blue stain (or ethidium bromide and UV source)

5 microcentrifuge tubes

0.5 - 10 ul. micropipettors and tips

37oC water bath

1 set electrophoresis equipment

1 microtube rack

10 mm rulers

10 sheets of graph paper

hot plate with magnetic stirrer or microwave oven

*please note that the above materials and a good portion of the lab can be cut out if one would rather
supply students with a printout of a gel that has already been run.

D. Estimated Time:
The time required for the lesson is 2-3 50 minute class periods if doing the actual gel electrophoresis. Without running the
gel it could be completed in a single 50 minute class period.

E. Procedure:
Teacher Preparation:
KEEP IN MIND THAT THE DNA SAMPLES IN THIS LAB ARE ACTUALLY DIFFERENT RESTRICTION ENZYMES AND THE
RESTRICTION ENZYME IS ACTUALLY LAMBDA DNA!!
Before class the teacher should prepare the DNA gel as well as aliquot the three different enzymes (labeled as DNA for the
demonstration), the DNA (labeled as enzyme), and the buffer solution. A 37C water bath should be prepared in a beaker.
Demonstration:
After the students have read the case file the teacher should now gather the students around the demonstration table. To
save time, tell the students that you are going to add an enzyme to the suspects DNA that will cut the DNA so that they
will be able to tell them apart. Add 4ul EcoRI (labeled suspect 1 DNA) to 4ul lambda DNA (labeled Restriction Enzyme) in a
1.5ml reaction tube and to that add 8ul 2X buffer. Do the same with the remaining two enzymes but only use 2ul of each
enzyme and 2ul DNA with 4ul 2X buffer. Incubate the 3 tubes for 45min in the water bath. After incubation, add 1ul
loading dye to each of the tubes. Next, pipette 10-15ul of each sample into the wells of the gel. In the first or last lane on
the gel, pipette 8ul of the 1Kb DNA ladder. Run electrophoresis at ~100v for about 30-45 minutes. During the time that
the gel is running is a good time to teach the students how to generate the first standard curve using the ladder that is
shown on the gel photo of the crime scene DNA. Also, go over how restriction enzymes act to cut DNA at specific sites
allowing for differentiation of DNA strands from different individuals. Explain how electrophoresis works because DNA is
negatively charged and will go toward the positive electrode and that DNA strands of different sizes travel different
distances. Smaller strands will travel shorter distances than larger fragments. After the gel has run and been stained.
Make a copy of it for each group so that each group can generate their own standard curve to determine the size of the
DNA fragments and compare that with the size they found the crime scene DNA to be. If using Caroline Blue stain to
stain the DNA bands, read the instructions that come with the kit.

F. Closure:
A great closure for this lesson is to display, read, or hand out a list of all of the people that have been exonerated and let
out of prison based on this procedure of DNA fingerprinting. Following this lesson plan is an up-to-date list including the
names and lengths of sentences served for crimes they did not commit.

G. Standards:
Science as Inquiry
CONTENT STANDARD A:
As a result of activities in grades 5-8, all students should develop:

Abilities necessary to do scientific inquiry

Understandings about scientific inquiry

Life Sciences
CONTENT STANDARD C:
As a result of their activities in grades 5-8, all students should develop understanding of:

Reproduction and heredity

Diversity and adaptations of organisms

Science and Technology

CONTENT STANDARD E:
As a result of activities in grades 5-8, all students should develop:

Abilities of technological design

Understandings about science and technology

H. Assessment:
Take up students reports and graphs and grade them on completion, content, writing skills, and neatness
Throughout the class period, walk around and assess participation
Give handout of similar who done it activities (possibly a good place to work in paternity tests)

I. References:
Web page to find national science teaching standards: http://www.nap.edu/readingroom/books/nses/html/
Web page to find supplies to teach biology: https://www3.carolina.com/onlinecatalog/
Web page with a list of individuals cleared by DNA evidence: http://www.pbs.org/wgbh/nova/sheppard/cleared.html

Cleared by DNA
The sheer number of inmates who have gained their freedom since the advent of forensic DNA
testing in the late 1980s may surprise you. Below is an up-to-date list of those freed inmates,
most of who were incarcerated for murder or rape. The list is maintained by the Innocence Project
at the Benjamin N. Cardozo School of Law in New York City.

Date of
Conviction

Date of
Release

Length of
Incarceration
(years)

IL
TX
MD
CA
PA
IL
TX
NY
NY
NY
NC
IL
NY
WV
WV
CA
IL
OK
OK
MD
DC
CA
CT
WV
IL

Oct-78
Oct-90
Mar-85
Oct-90
Jun-91
May-84
Aug-85
Mar-87
Jun-87
Aug-85
Jan-85
Jan-85
Apr-84
Jan-87
Jul-86
Aug-84
Jul-79
Mar-93
Apr-88
Oct-91
Jul-89
Oct-80
Mar-90
Oct-87
Jan-85

Jul-96
Jul-94
Jun-93
Jan-94
Jan-94
Oct-94
Oct-97
Oct-92
Apr-95
Sep-87
Jun-95
Nov-95
Aug-91
Jun-94
Jul-94
Sep-94
Aug-89
Jan-97
Apr-99
Feb-99
Mar-90
Jun-96
Oct-92
Oct-95
Nov-95

19
14
8
4
4
10
12
6
7.5
4
11
11
9
7.5
8
10
8
5
12
7
0.75
16
2.5
8
11

WI
WV
VA
NY
IL
KS
GA
IL
IL
NY
IL
AL
AL
OK
MA
SC

Dec-91
Mar-84
Feb-85
Nov-83
Dec-85
Feb-86
Nov-83
Jan-92
Jul-89
Mar-82
Jun-82
Jun-84
Jun-84
Jan-88
Jan-90
Jan-84

Jul-96
Jun-99
Oct-94
not yet
Jun-96
Jul-92
Jun-99
Dec-95
May-99
Dec-92
Jan-85
Dec-97
Dec-97
Jan-98
Apr-97
Aug-98

5
15
10
U
11
7
16
4
10
11
3
14
14
9
8
14

Name of Defendant Location

Kenneth Adams
Gilbert Alejandro
Kirk Bloodsworth
Mark Diaz Bravo
Dale Brison
Ronnie Bullock
Kevin Byrd
Leonard Callace
Terry Chalmers
Marion Coakley
Ronald Cotton
Rolando Cruz
Charles Dabbs
Dewey Davis
Gerald Davis
Frederick Daye
Gary Dotson
Timothy Durham
Dennis Fritz
Anthony Gray
Edward Green
Kevin Green
Ricky Hammond
William Harris
Alejandro
Hernandez
Anthony Hicks
Larry Holdren
Edward Honaker
Vincent Jenkins
Verneal Jimerson
Jones Joe
Calvin Johnson
Richard Johnson
Ronald Jones
Kerry Kotler
Steven Linscott
Dale Mahan
Ronnie Mahan
Robert Miller
Marvin Mitchell
Perry Mitchell

Vincent Moto
Bruce Nelson
Victor Ortiz
Brian Piszczek
Willie Rainge
Donald Reynolds
Fredric Saecker
Ben Salazar
Dwayne Scruggs
David Shepard
Walter Smith
Walter Snyder
Steven Toney
David Vasquez
Billy Wardell
Thomas Webb
Troy Webb
Dennis Williams
Ron Williamson
John Willis
Glen Woodall
Herman Kaglick
David Milgaard
Gordon Folland
Guy Paul Morin
Gregory Parsons

PA
PA
NY
OH
IL
IL
WI
TX
IN
NJ
OH
VA
MO
VA
IL
OK
VA
IL
OK
IL
WV
CAN
CAN
CAN
CAN
CAN

Jan-88
Apr-85
Jan-84
Jun-91
Oct-78
Feb-88
Jan-90
Jan-92
May-86
Sep-84
Dec-86
Jun-86
Apr-83
Feb-85
Feb-88
Jan-83
Apr-89
Oct-78
Apr-88
Sep-90
Jun-87
Jan-92
Jan-70
Mar-95
Jul-92
Feb-94

Nov-95
May-91
Oct-96
Sep-94
Jul-96
Nov-97
May-96
Oct-97
Dec-93
Apr-95
Dec-96
Apr-93
Jul-96
Jan-89
Nov-97
May-96
Oct-96
Jul-96
Apr-99
Feb-99
May-92
Jan-98
Apr-92
Nov-96
Jan-95
Feb-98

9
9
14
4
18
9
6
5
8
11
10
7
14
5
9
14
7
18
12
9
4
5
23
3
2
0.25