ACTIVITY # 10

DNA Fingerprinting
GROUP 2- GONZALES, EYORCADAS, DABALOS, CULLAMAT

DNA is composed of four different chemical building blocks called "bases". These
four bases; adenine (A), guanine (G), thymine (T), and cytosine (C); are joined together
in a one strand by covalent bonds. These two strands are held together in a double helix
because bases with complementary shapes can pair with each other. Adenine is able to
pair with thymine and guanine pairs with cytosine.
Each human cell contains approximately 3 billion base pairs of DNA organized in 23 pairs
of chromosomes. The two sets of chromosomes from the mother and the father are very
similar. For example, the sequence of the bases in chromosome 1 is almost the same in
every human; however, there are specific regions in the chromosomes where the sequence
can vary, on the average one base out of every 700-1000 bases will differ between two
individuals.

Techniques used for DNA fingerprinting

DNA is cut into different pieces by enzymes that recognize specific DNA
sequences, the sizes of these fragments can be measured and will vary between individuals,
each pattern of fragments occur with a certain frequency in a population, and we can
calculate the likelihood of finding a specific combination of patterns within a population.
A hypothetical example of a VNTR found in humans is a chromosomal site known
as H1Z38. The size of the repeating sequence in H1Z38 is 4 base pairs long. The H1Z38
sequence is located on chromosome 1 and is usually repeated between 2 and 40 times.
Restriction enzymes are used to cut DNA on both sides of the repeating sequence,
generating a fragment whose length is determined by the number of repeats as shown
below.

5’ GAATTCCCATCCATCCATCCATGAATTC 3’
3’ CTTAAGGGTAGGTAGGTAGGTACTTAAG 5’

5’ AATTCCCATCCATCCATCCATG 3’
3’ GGGTAGGTAGGTAGGTACTTAA 5’

Objectives:
At the end of the session, the students are expected to:

1. paraphrase the steps in DNA fingerprinting;

2. compare RFLP among individuals;
3. analyze RFLP patterns; and
4. calculate the likelihood of finding a specific combination of patterns within a
population.
Materials:
Paper strips with sequences of DNA bases
Scissors
Manila paper
Colored pentel pen
Paste
Masking tape

Procedure:

Read the situation below under the “The crime”.

1. On a piece of paper with a printed DNA sequence, look for the following
sequence: 5' GAATTC 3' (the 5' and 3'numbers show the orientation of the DNA
sequence); when you find this sequence, cut the sequence after the "G".
2. Count the number of 5’ CAATT 3’ repeats on each fragment.
3. After counting the number of 5’ CAATT 3’ repeats on each fragment, tally the
results in a table on the board (table 1).
4. Draw a diagram of an agarose gel on manila paper (figure 1).
5. Paste fragments on agarose gel diagram on the lane of the suspect according to
fragment size. Smaller fragments travel faster and thus, are relatively farther from
the well.
6. Compare the pattern of restriction fragments from blood found at the crime scene
to the patterns from the potential suspects and the victim.
7. Calculate the probability of finding the culprit’s DNA fingerprint in the general
population. The probability of finding any one pattern is equal to the number of
times that a specific pattern is found divided by the sample size. The number of
students holding the “General population” sequences will determine the sample
size. The probability of finding a combination of two or more restriction patterns
is determined by multiplying the probability of finding each pattern, this
relationship is also known as the product rule.

The crime
Paul’s body was found at midnight in the university library by the janitor. A
computer monitor lay near the body in a pool of blood. The suspects include Kimor, Annie
and Poley. Kimor, a wealthy middle-aged heir, was known to hate Paul for stealing the
attentions of his ex-gf, Daenerys. Annie, the notorious con artist, may have had a reason
to silence Paul, because only she knew her true identity. Perhaps even sweet Poley had her
own reasons. She despised Paul and blamed him for breaking the heart of her only
daughter, which eventually led to her death.

Murnee’s body was found at midnight in the university library by the janitor. A
computer monitor lay near the body in a pool of blood. The suspects include Ditdit, Daren,
and Jem. Ditdit, a wealthy middle-aged heiress, was known to hate Murnee for stealing
the attentions of her ex-suitor, Sam. Daren, the notorious con artist, may have had a reason
to silence Murnee, because only she knew his true identity. Perhaps even sweet Jem had
her own reasons. She despised Murnee and blamed Murnee for breaking the heart of her
only son, which eventually led to his death.

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DN Lane 1 Lane 2 Lane 3 Lane 4 Lane 5
A
size
Crime Scene Paul Kimor Annie Poley
Murnee Ditdit Daren Jem

11 5’ GAATTC
GAATTC 3’

5’ GAATTC
GAATTC 3’

5’ GAATTC
GAATTC 3’

5’ GAATTC
GAATTC 3’

5’ GAATTC
GAATTC 3’

5’ GAATTC
GAATTC 3’

6 5’ GAATTC 5’ GAATTC
GAATTC 3’ GAATTC 3’

5’ GAATTC 5’ GAATTC
GAATTC 3’ GAATTC 3’

5’ GAATTC 5’ GAATTC
GAATTC 3’ GAATTC 3’

4 5’ GAATTC 5’ GAATTC
GAATTC 3’ GAATTC 3’

5’ GAATTC 5’ GAATTC
GAATTC 3’ GAATTC 3’

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Figure 1. Agarose gel

Fragment size Allele frequency

1 1/60
2 2/60
3 1/60
4 1/60
5 2/60
6 1/60
7 1/60
8 1/60
9 1/60
10 1/60
11 1/60

CRIME SCENE
5’ GAATTCCCATCCATCCATCCATCCATCCATCCATCCATCCATCCATGAATTC 3’
3’ CTTAAGGGTAGGTAGGTAGGTAGGTAGGTAGGTAGGTAGGTAGGTACTTAAG 5’

5’ GAATTCCCATCCATCCATCCATCCATCCATCCATCCATGAATTC 3’
3’ CTTAAGGGTAGGTAGGTAGGTAGGTAGGTAGGTAGGTACTTAAG 5’

5’ GAATTCCCATCCATCCATCCATCCATCCATGAATTC 3’
3’ CTTAAGGGTAGGTAGGTAGGTAGGTAGGTACTTAAG 5’

5’ GAATTCCCATCCATCCATCCATCCATGAATTC 3’
3’ CTTAAGGGTAGGTAGGTAGGTAGGTACTTAAG 5’

5’ GAATTCCCATCCATCCATCCATGAATTC 3’
3’ CTTAAGGGTAGGTAGGTAGGTACTTAAG 5’

5’ GAATTCCCATCCATGAATTC 3’
3’ CTTAAGGGTAGGTACTTAAG 5’

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 Paul/Murnee
5’ GAATTCCCATCCATCCATCCATCCATCCATGAATTC 3’
3’ CTTAAGGGTAGGTAGGTAGGTAGGTAGGTACTTAAG 5’

5’ GAATTCCCATCCATCCATCCATGAATTC 3’
3’ CTTAAGGGTAGGTAGGTAGGTACTTAAG 5’

5’ GAATTCCCATCCATGAATTC 3’
3’ CTTAAGGGTAGGTACTTAAG 5’

Kimor/Ditdit
5’ GAATTCCCATCCATCCATCCATCCATCCATCCATCCATCCATCCATCCATGAATTC 3’
3’ CTTAAGGGTAGGTAGGTAGGTAGGTAGGTAGGTAGGTAGGTAGGTAGGTACTTAAG 5’

5’ GAATTCCCATCCATCCATGAATTC 3’
3’ CTTAAGGGTAGGTAGGTACTTAAG 5’

Annie/Daren
5’ GAATTCCCATCCATCCATCCATCCATCCATCCATCCATCCATCCATGAATTC 3’
3’ CTTAAGGGTAGGTAGGTAGGTAGGTAGGTAGGTAGGTAGGTAGGTACTTAAG 5’

5’ GAATTCCCATCCATCCATCCATCCATCCATCCATCCATGAATTC 3’
3’ CTTAAGGGTAGGTAGGTAGGTAGGTAGGTAGGTAGGTACTTAAG 5’

5’ GAATTCCCATCCATCCATCCATCCATGAATTC 3’
3’ CTTAAGGGTAGGTAGGTAGGTAGGTACTTAAG 5’

Poley/Jem
5’ GAATTCCCATCCATCCATCCATCCATCCATCCATCCATCCATGAATTC 3’
3’ CTTAAGGGTAGGTAGGTAGGTAGGTAGGTAGGTAGGTAGGTACTTAAG 5’

5’ GAATTCCCATCCATCCATCCATCCATCCATCCATGAATTC 3’
3’ CTTAAGGGTAGGTAGGTAGGTAGGTAGGTAGGTACTTAAG 5’

Questions:

1. How many suspects are guilty? Who are guilty? Why?

There are 2 suspects guilty in the crime. Annie is the suspect for the crime of Paul’s
body, while Daren is the suspect for the crime of Murnee’s body. Their DNA sequence
found is identical to the DNA sequence of Paul and Murnee found in the crime scene.
Thus, we are able to identify which are the suspect among them in the crime scene
according to their DNA sequence.

2. Based on the DNA evidence, how can you say that a suspect is innocent?

No two people, with the exception of identical twins, have the same DNA, making it
an effective investigative tool. As a result, DNA evidence collected at a crime scene
can be used to identify a suspect, establish their innocence, or rule them out of further
investigation. Therefore, if the DNA of the suspect does not match the DNA at the
crime scene, the suspect is innocent.

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 3. What is the probability that the culprit is from the general population and not
among the suspects?

Documentation: