BIOLOGY 3

Philippine Science High School – Main Campus

Activity on DNA Fingerprinting

Background on DNA Fingerprinting

The term "fingerprinting" is used to indicate that each individual can be

identified by the unique pattern that exists on the fingers of that individual. No two
people have identical patterns. This is also true of an individual's DNA. No two
people have the same DNA. Researchers can isolate DNA from an individual and
create a "fingerprint" of that individual.

This technology has many applications. Suspects have been convicted, as

well as acquitted, of crimes based on evidence found at the scene of the crime, such
as a drop of blood. Paternity cases have been argued using this technology: the DNA
of the child is compared with that of the possible father to see if the child has any of
the same patterns of DNA, indicating a biological relationship. Individuals can also be
tested for genetic diseases that run in a family.

DNA fingerprinting relies on the ability of specific enzymes to cut the DNA into
small pieces and the ability to analyze those pieces and compare them between
samples. The DNA is cut by a type of enzyme called a restriction enzyme.
Restriction enzymes occur naturally in bacterial cells as a line of defense, similar to
our immune systems. The enzymes are very specific: each will recognize only a
certain site on a DNA molecule and cut only at that site. These sites occur randomly
anywhere on the DNA molecule. Therefore, every different DNA molecule (for
example, DNA from two different people) will be cut into a different number of pieces
and pieces of different sizes when incubated with a restriction enzyme. In addition,
no two restriction enzymes will cut the same DNA molecule in the same way.
Therefore, no two enzymes used to cut the same DNA molecule will give you the
same number of pieces or the same sized pieces of DNA.

After the DNA is cut, it is necessary to determine exactly how the enzymes
cut the DNA and to measure the size of the pieces of DNA that have been cut. This
analysis is done by a process called gel electrophoresis. Electrophoresis is possible
because DNA has a negative charge. Therefore, if an electric current is applied to the
DNA, the DNA will move towards the positive pole. However, moving the DNA in this
way will not separate the DNA pieces based on their sizes. That's where the "gel" in
"gel electrophoresis" comes in. The gel is made of agarose, a substance that forms
pores that the DNA has to move through. The larger the size piece of DNA, the more
difficulty it will have moving through the pores in the gel and the slower it will move.
In this way different sized pieces of DNA can be separated, and pieces that are the
same size will move to the same position in the gel.

After electrophoresis, the pieces of DNA will appear as bands in the gel. Each
band contains thousands of DNA molecules, which are all the same size. This array
of bands forms the pattern that is called a "fingerprint." It is also necessary to run a
marker of known size on the gel. The size of each piece of DNA can be determined
based on how it moved in relation to the marker.

There is of course a random probability that anyone’s DNA could match

someone else’s at one point (called a locus). But the chance that the DNA of two
different people will match exactly at four or five or six different points (loci) is highly
improbable. You may have heard of statistics in the range of one in a billion as the
chance two random samples could match at all loci.
 Though DNA fingerprinting has proven have various applications, there are
also controversies surrounding this technology. Some argue that the techniques used
are not fool proof and therefore they should not be used to convict someone of a
crime. Some people worry about who should have access to the information obtained
from individuals tested for genetic diseases. For instance, many believe insurance
companies would deny coverage for people who are determined to be predisposed to
diseases based on information gained from DNA fingerprinting. Other types of
discrimination may occur based on this information. For instance, an airline may not
hire a pilot if that person was found to have a gene causing a predisposition to high
blood pressure. This person may be more likely than most people of having a heart
attack, and this would endanger many lives if the person were piloting a plane when
and if a heart attack occurred.

In this activity, you will simulate what is done when DNA fingerprinting is used
as forensic evidence in a given crime.

Materials
1. Different pieces of colored paper will be given to each group. This will
represent different DNA samples gathered from suspects and the crime scene.
2. Each group will also be assigned a restriction enzyme and its recognition
sequence. (NOTE: In reality, 4 or more different restriction enzymes are used in
combination to produce a more unique fingerprint. But to simplify this simulation,
only one restriction enzyme will be used by each group.)
3. The piece of manila paper will act as the gel where the fragments produced
by the restriction enzymes will be separated.

Procedure
You have been tasked to analyze the DNA from the following case:
Three robbers broke into an electronics shop late one night and stole equipment
and cash. They were careful to wear gloves and not leave any traditional
fingerprints. However, one of them was cut when climbing through the broken
window, and a small sample of blood was left behind. Several days later, the police
picked up three men selling stolen electronic equipment and believed them to be the
ones who robbed the shop. Samples of blood were taken from each suspect and the
DNA was isolated. Genetic material was also isolated from the blood sample left at
the crime scene.

1. Cut each DNA sample into fragments by searching for the restriction sites of
the given restriction enzyme.
2. Paste the various fragments according to size (base pairs or “bp”) on the gel
(manila paper) by following the marker on the left.
3. Compare the band patterns produced by the fragments from different DNA
samples? Does the pattern of bands from the blood at the crime scene look
EXACTLY like the pattern of bands of one of the suspects?
4. Based on your observations in number 3, decide whether you can prove that
one of the 3 suspects was at the scene of the crime.

Questions for further thought:

1. What are some other possible uses of this technology? What are some other
benefits and problems associated with DNA fingerprinting?
2. If a genetic disease were carried in your family, would you want to be tested
for it? Would you want your unborn child tested for it?
3. Who should be given access to information gained from DNA fingerprinting?
4. There are many issues involved in this topic. What further information do you
think you need in order to continue discussing these questions and to make
informed decisions about this technology?