Name: Mike Savio

Biology Period ____________

Date _______________________

DNA Fingerprinting Lab – Whodunnit? (EDVOTEK 130)

Purpose: The purpose of this activity is to use gel electrophoresis to analyze the DNA found at a crime
scene and from three suspects.

Pre-lab Assignment:

Read the attached “Background Information” AND the procedure. Then answer the questions below. Some
questions are related to information learned earlier this year.

1. Name three situations in which you can use DNA fingerprinting.

Some uses could include, Determining the father of a baby, identifying genetic diseases, and identifying
human remains.

2. What is the purpose of the enzyme in this lab? Be as detailed as possible.

The enzyme cuts the blood sample and the suspect’s DNA.

3. What will you load into each well of the gel?

DNA samples will be loaded into the wells of gel.

4. Which color cord gets connected to the – electrode?

The black cord is connected to the - electrode.

5. Which color cord gets connected to the + electrode?

The red cord is connected to the + electrode.

6. What charge does DNA have? What charge is DNA attracted to?
DNA has a negative charge. DNA is attracted to positive.

7. How long will you run the gel for?

The gel will run for 30 min – 1 hr.

8. How long will you stain the gel for?

The gel will stain for 5 minutes.

9. How long will you destain the gel for?

The gel will be destained for 20 minutes.

Procedure:
Prepare the Gel
(Materials: Agarose, buffer, distilled water, scale, hot plate, flask, stirring rod)

0.39 g of agarose + 1.0 mL 50x conc. Buffer + 49.0 mL d. H20 (heat while stirring until clear)

Load the Samples and Run the Gel

(Materials: gel box, gel, power source and cords, 500 mL buffer, DNA samples, p200 micropipettes)

1. Place gel into the gel box.

2. Add buffer into the gel box until the gel is submerged.
3. Tap each tube before loading to ensure that all of the sample is at the bottom of the tube.
4. Load each well with 35µL of DNA according to the following guide:

Lane Tube
1 A DNA Standard Marker 1
2 B Crime Scene DNA cut with enzyme
3 C Suspect 1 DNA cut with enzyme
4 D Suspect 2 DNA cut with enzyme
5 E Suspect 3 DNA cut with enzyme

5. After DNA samples have been loaded, connect the gel box to power source.
Remember black is –
Red is +

6. Set the power source to the required voltage – 125 V.

10. Check to see that the electric current is flowing properly. You should see bubbles forming on the two
electrodes. Let the gel run for the length of time specified by your teacher (30 min – 1 hr.)

Stain the Gel

(Materials: per gel: 75 mL of 1x FlashBlue Stain, 300 mL d. H20, 2 trays gloves, goggles, funnel

1. Carefully remove the agarose gel from its bed and completely submerge the gel in a small tray
containing 75 – 100 mL of 1x FlashBlue Stain. Add more stain if needed to completely submerge the
gel.

2. Stain the gel for 5 minutes.

3. Carefully transfer the gel to another small tray and fill it with 250 – 300 mL of distilled water.

4. Gently agitate the tray every few minutes for 20 minutes to destain the gel. Dark blue bands will
become visible against a light blue background. Additional destaining may be required to yield
optimal results.

Results
Draw your results in the diagram below. Then answer the questions that follow.

1 2 3 4 5 6
________ ________ ________ _______ ________ ________

1. Which suspect’s DNA was found at the crime scene? Explain how you know.

2. Name and explain two sources of error in this lab.

3. Explain a procedure that could be used to determine which of 5 suspects got into a violent bloody fight
and then murdered a store owner. As part of your answer be sure to explain what you will put in each
well of the gel.

Background Information
DNA fingerprinting is a technique used to analyze DNA samples by cutting the DNA with an enzyme and
analyzing the various size fragments created. The technique works because within each person’s DNA, there
are differences. Dr. Alex Jeffreys first discovered DNA fingerprinting at the University of Leicester in 1984.
It was used to solve a murder for the first time in 1987. Then, two months later, it solved the first case in the
U.S. Since then, DNA fingerprinting has been used in thousands of cases – both to convict suspects and to
free those who have been wrongly convicted. Other uses for DNA fingerprinting include determining the
father of a baby, identifying genetic diseases, and identifying human remains from war casualties or other
horrific events such as the bodies of the victims of 9-11.

The reason why DNA fingerprinting works is because each person’s DNA is slightly different and will be cut
by enzymes at different spots. Each person’s DNA will create different number of fragments and different
size fragments. These fragments will travel through the gel at different speeds creating a unique “DNA
fingerprint” for each person. In forensic cases, DNA samples can be obtained from small bits of skin, blood,
semen, or hair roots. The DNA fingerprint created from these samples can be compared to the DNA
fingerprints created from the suspects’ DNA

In this lab, you will use DNA fingerprinting to determine who broke into the school a few Saturdays ago.
Three weeks ago, the school was broken into late on a Saturday night. The glass in the front door was
broken, the entire front hallway was vandalized, and the alarm was set off. When police arrived to
investigate the alarm, they noticed small bits of fiber from a shirt as well as blood on the broken glass of the
door. After questioning various students, the suspects were narrowed down to three. A blood sample was
taken from each suspect. Each suspects’ DNA was cut with an enzyme. The crime scene blood was also cut
with the same enzyme. By analyzing the DNA fingerprint, you will determine which suspect committed this
crime.