DNA FINGERPRINTING

- Discovered that variable number tandem repeats

BSMT 2 | S.Y. 2021-2022 Second Semester – FINALS (VNTR) - a part of junk DNA - vary from individual to
another
CYTOGENETICS LESSON # 12
DNA FINGERPRINTING
Lecturer: Mr. Nino Paolo Tan

Topic Outline
● What is DNA fingerprinting?
● Who discovered it?
● How is it done?
● Why do we do it?
● For whom is it?
• For his study, he tried looking into the DNA. He
discovered that there is something that we call as
WHAT IS DNA FINGERPRINTING? VNTR.
● A molecular method for identifying an individual member • This VNTR is a part of junk DNA meaning it is a
of a population or species. noncoding DNA and it varies from one individual to
● Individual identification by means of differences in DNA another. So just by using this junk DNA, there is now
sequence is generally visualized as the pattern of DNA variation between individuals and we can use that.
fragments after separation by gel electrophoresis.
● With technological advancements in modern methods, • It was first used to help resolve an immigration case in
now more commonly called DNA profiling. 1985.
EXPLANATION: • First applied in forensic science in 1986 to analyze
• The use of molecular genetic methods to determine the semen samples recovered from two rape victims.
exact genotype of a DNA sample and in a way, this can • In 1987, Colin Pitchfork became the first person to be
basically distinguish 1 human being or individual from identified and convicted as a result of DNA
another just by using that DNA sample fingerprinting.
• By looking at the differences in the DNA sequence, there
is a pattern and that pattern is unique to that individual.
This is now more commonly called DNA profiling.
• To give you a comparison - the normal fingerprinting that
you know of is when we use our actual fingers. In this
fingerprinting the pattern is formed by the unique ridges
that are found in our fingers.
• The ridges or lines form a unique pattern and that pattern
serves as an identifier for that person.
In just a short time, results of DNA fingerprinting are
• The same thing happens for DNA fingerprinting but for evident. It is also used in other fields because Dr. Alec
this one the pattern is formed by the differences in the Jeffreys was just working on immigration disputes,
DNA sequence. hereditary diseases and this time the application for his
• Since each individual has a different DNA sequence from work is on forensic science.
each other, the pattern that is now formed by the
differences in our DNA sequences serves as the
identifier.
• We can identify one person from the other because of the
unique pattern formed by the DNA sequence just like in
the actual fingerprinting. One person can be identified
because of the unique pattern formed by the lines or
ridges of the fingers.

WHO DISCOVERED DNA FINGERPRINTING?

● Discovered and developed by Dr. Alec Jeffreys in 1985
at the University of Leicester in England
- In 1984, Dr. Jeffreys was studying hereditary
diseases in families and also developing methods to
resolve paternity and immigration disputes by linking
genetic elements between individuals.
KENNY. LOPEZ. PANOREL. PISCADERO. ROLLENAS. SANICO. SO. SULAPAS. TAMAYO. VERGARA. VILLAROYA
In this picture is DNA. So, there are exons and introns.
We all know that introns are our noncoding DNA, they are
the junk DNA.
Question: Unique mn jd na atong DNA from one person
to another but why are we focusing on the noncoding
DNA or the junk DNA?
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• Scientist and those in the forensic community, they

prefer to use junk DNA because of the very fact that
they are noncoding, they don't produce a trait.
• So, it is safer for the privacy of the individual. They
do not carry any information about gene expression
of the patient or person.
• They only provide information about relatedness and
they don't show any about a person’s biological state
(e.g., physical appearance, and even their mental
wellbeing).
• Because in the end we are just looking for
relatedness, if the DNA of this person is related to the
DNA that we found in the crime scene. We don't need
• We are just looking at the patterns that are being
formed but not by the ridges of our fingers but by the
to know what that person really looks like.
variations in the number of times that each sequence
● Satellite - short segment of repeated DNA sequences (if was repeated.
mag repeat gani ang DNA sequence satellite na siya) • Again, it is not the variation sa sequences because
● Minisatellite - 10-100 base pairs of nucleotides (10-100 the same man ang sequence nga atong gi look into
ang magsige og repeat) but what matters is the number of times that, that
● Microsatellite - 2-10 base pairs of nucleotides (2-10 ang specific sequence was seen in an individual. So mao
mag sige og repeat) na siya ang DNA fingerprinting.
(These 2 are the ones that we primarily look into when we are
doing DNA fingerprinting) HOW IS DNA FINGERPRINTING DONE?
For this discussion we will just talk about 4 methods:
● VNTR-based DNA fingerprinting
● RFLP-based DNA fingerprinting
● STR DNA profiling
● Next Generation Sequencing DNA Profiling

1. VNTR-based DNA Fingerprinting (Variable Number

Tandem Repeats)
• For this method we have 4 steps:
To amplify specific DNA
Polymerase Chain Reaction
fragments
To separate the
Gel Electrophoresis
fragments
Southern Blot Technique To detect by blotting
Here, we have 2 chromosomes, representing 2 To transfer results in a
individuals labeled person A and person B. When we look Visualization
permanent copy
into the DNA sequence these are found at the telomere (DNA fingerprinting is an application of the techniques that we
near the end of the chromosome. have learned so far)
• First, in the minisatellite, naa siya ana nga
sequence, it's a bit longer but it falls in the 10-100 • Made up of short segment DNA sequences/Satellites that
base pairs of nucleotides. TTAGAGATCCAGGG are between 10 and 100 bp long (Minisatellites), with
mao na siya ang first sequence. each unit repeated two or several hundred times. (VNTR
• Now, person A it is only seen once but person B, the and Minisatellites mean the same. In VNTR we are
pattern is repeated twice. So just by looking at that looking for VNTRs or Minisatellites not Microsatellites.)
pattern, we can identify person A from person B. • Often clustered near the telomere or end of the
• They have exactly the same sequence but the chromosome.
variation comes from the number of times na gi • These repeat sequences can vary from person to person.
repeat ang sequence.
• Now microsatellites, much shorter. So ang gi repeat 1st Step: Polymerase Chain Reaction
is CA. So person A CACACACACA so na repeat 5 ● We do this to make enough copies of DNA to perform the
times while in person B exactly the same sequence analysis (We need to have lots of copies of DNA mn jd to
but seen 7 times. have a stronger reaction and to better visualize this
reaction)
• Again, the same sequence but what varies is the
If naa natay millions of copies of DNA that we want, we do a
number of times na repeat. 5 times sa person A nya
separation technique.
7 times sa person B. So naa nay variations then we
can use that to differentiate person A from person B
and therefore identify them.

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2nd Step: Gel Electrophoresis
● DNA fragments are separated by size (length, or
molecular weight by gel electrophoresis) according to
their charge
● Denatured by alkali (NAOH) to produce Single-Stranded
fragments (after this step all the DNA present are now
single stranded)
Visual representation of what happens during the gel
electrophoresis step. In gel electrophoresis we separate the
different DNA fragments according to their molecular weight
or their length or how heavy they are.
KENNY. LOPEZ. PANOREL. PISCADERO. ROLLENAS. ROMA. SANICO. SO. SULAPAS. TAMAYO. VERGARA. VILLAROYA
• For sample A has the corresponding fragments. For
DNA fingerprinting they each have the same
sequence or the same fragments but what differs is
the number of times that they are seen or repeated.
• We can see nga si sample A has the same sequence
but is seen twice while si B the same sequence but
seen three times and si C the same fragments but it
is seen five times.
• Si C ang longest and also the heaviest while si A
much more shorter kay ka 2 times raman siya ma
seen so it is the lightest.
• So, if you do gel electrophoresis A will travel the
furthest distance while si C since a bit longer and
heavier siya than the other samples si C only traveled
the least distance. So that's what happens in gel
electrophoresis. That's how we can separate the
different DNA samples that we have.
3rd Step: Southern Blot Technique
(In this step we are now actually partially visualizing our
results)
● Probes loaded with radioactively labeled DNA are added
● These probes bind to the DNA fragments with
complementary sequences making them double-
stranded (Because dba in the previous step all DNA
fragments now are single stranded. So, when this step
occurs the radioactively labeled DNA will bind with those
fragments that they are complementary into and make
them double stranded again. While those fragments nga
not complementary with our radioactively labeled DNA,
they will remain as single stranded)
● Unbound fragments are washed out (wash out procedure
in which the remaining unbound fragments, the remaining
single stranded fragments are washed out the solution at
the end of this step. (shawawt mga single :/ wakakaka)
(If by this step wala sila ka bind og radioactively labeled DNA
and wala sila nabalik into double stranded then they are
washed out in the solution. The only thing that remains here
are the double stranded DNA that are the very ones that we
want to test on.)
Over here the fragments are visualized by Southern Blotting:
the separated fragments are transferred from the gel to nylon
paper. Then a radioactively labeled DNA probe is added. The
probe binds to those DNA fragments with complementary
sequences.
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This is an example of an autoradiograph. So, this one is

used in DNA fingerprinting and this is used for a forensics
case.
o D – defendant (murag suspect sa case)
o Jeans – jeans of defendant
o Shirt – shirt of defendant
o Black thingy – DNA fragment
o V – Victim (under niya is DNA fragment sa victim)
• If you look closely lahi jd ang pattern sa defendant
and the victim because they are different people.
• TS is the control and we also have what we call as
the ladders and are placed at both ends so that it is
easier for you to visualize or compare.
(It is like this, so si Southern blot technique first imo siya • So unsa mn ang case dinhi, we already know nga si
i.incubate in probe solution then itransfer nimo ang results into defendant lahi jd iyang DNA sa victim but the jeans
a piece of nylon sheet or nylon paper. We can now partially and shirt sa defendant contains DNA from the victim.
visualize the results. Pero a bit harder to see since wala pa • So, from this result, you should already know na
tay final visualization step) since si defendant mn ang gawear ani normal ra nga
naay DNA from the defendant na ma found sa jeans
4th Step: Visualization and shirt but it is suspicious if naay DNA from the
Last step in VNTR-based DNA fingerprinting. We don't want victim that will be found on the shirt of the defendant.
our results to remain sa nylon paper or nylon membrane, • What does this mean? Does this mean nga ang
because ang paper dali ra ma damage, mabasa, magisi. So, defendant is the suspect? Not yet, we cannot really
for this one, we want to transfer it to another form of hardcopy, claim that pa but basing on the results then we can
we use film. say that potentially the defendant was in the
● The blot is covered with radiation-sensitive film to give an presence of the victim because of the DNA results
autoradiograph. This shows the location of those DNA nga ang DNA sa victim was found sa shirt sa
fragments that reacted with radioactive probe. (So, the defendant.
same results but now much better lang ang hard copy) • So kani siya when you are doing DNA fingerprinting,
● The final product of a DNA fingerprint is an you are not part of the police or the investigating body
autoradiograph that contains at least five essential lanes. gyd nga maka claim na gyd using this one evidence.
● The markers are standardized DNA fragments of known This is just one of the many evidence that can be
size, which have been radioactively labeled. They help used to further make a strong case.
determine the size of the various fragments.
● The “control” is DNA from a source known to react
2. RFLP-based DNA Fingerprinting (Restriction
positively and reliably to the DNA probes and shows
Fragment Length Polymorphism)
whether the test has worked as expected.
• 4 Steps:
○ So, five essential lanes man siya, one of the
DNA digestion To cut DNA fragments
lanes is your control because dba when you do
testing you should always have a control. So Gel Electrophoresis To separate the fragments
kaning control contains known DNA.) Southern Blot Technique To detect by blotting
To transfer results in a
Visualization
permanent copy
(As we can see different ra siya sa first step instead of the
PCR step we have our DNA digestion step in which we use
DNA restriction enzymes to cut up DNA fragments so that is
the first difference between RFLP and VNTR. But they have
exactly the same, even down to the minisatellites or VNTR,
RFLP-based is a kind of VNTR-based DNA fingerprinting in
that we are still using the minisatellites or the VNTRs as our
basis for the differentiation when we do DNA fingerprinting.)

Actual DNA fingerprinting showing that the pattern of DNA

fragments of the victim (V) were found on the defendant’s
clothing(jeans/shirt). The first two and last two lanes are the
standard size markers (labeled #lambda and 1 kb). The lane
marked TS is a positive control showing that the fingerprint
technique was successful. The lane marked D is the
defendant’s DNA pattern.

KENNY. LOPEZ. PANOREL. PISCADERO. ROLLENAS. ROMA. SANICO. SO. SULAPAS. TAMAYO. VERGARA. VILLAROYA 4
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1st Step: DNA Digestion
● First DNA fingerprinting procedures are RFLP-based.
● Non-PCR based - uses restriction enzymes to cut DNA
fragments.
● Various restriction enzymes with distinct recognition sites
exist (in practice, several are used in each DNA sample)
(This step is a very old method, this was the first DNA
fingerprinting procedure, this was made before the discovery
of PCR so that is why wala pay PCR step because wala pay
PCR when this was made. What we do is that we use
restriction enzymes to cut up the DNA fragments that they
want to test on. There are various restriction enzymes with
their distinct recognition sites.)
• In A in sample 1, it is repeated significantly less
amount of times compared to Sample 2, which is
longer.
• It will also translate in the electrophoresis step. Since
A of Sample 1 is much shorter, it was able to travel a
much farther distance than Sample 2, which is very
long and was significantly repeated more times, so
distance traveled is very short.
• That is the principle and how we separate fragments
in electrophoresis.
RECAP!
● VNTR-based DNA fingerprinting uses the PCR
step as the first step in DNA fingerprinting
● RFLP-based DNA fingerprinting was made before
the discovery of PCR, so it has the DNA digestion
step instead
● BOTH uses minisatellites or VNTRs as their basis for
their variation for their DNA Fingerprinting

3. STR-based DNA Fingerprinting (Short Tandem

Repeats)
(we have here a table ‘no need to memorize’. In here are the Polymerase Chain To amplify specific DNA
restriction enzymes and the role of restriction enzymes. Each Reaction fragments
restriction enzymes kay naay name and mostly they come
from our bacteria, and even sa strain very specific siya, and of Gel Electrophoresis To separate the fragments
course each restriction enzymes they have their own unique
recognition site.) Southern Blot Technique To detect by blotting

Visualization To transfer results in a

● These sequence differences are called RFLP.
● Even if mutations have changed a few bases of the target
sequence, there is still enough similarity for probes to
bind.
● Variations in the DNA base sequence of restriction
enzyme recognition sites result in differences in the
fragments’ sizes.
(After the DNA digestion step, after na cut up into numerous
fragments, they are different sizes. After this you can produce
fragments nga different sizes.)
permanent copy
Steps are exactly the same as the first method discussed,
VNTR-based DNA fingerprinting. What differs here is that
instead of using minisatellites, microsatellites are used.
Microsatellites are much shorter (2 to 10 base pairs).
REMEMBER!
VNTR and RFLP based DNA fingerprinting uses
MINISATELLITES, whereas, STR (short tandem repeats)
uses MICROSATELLITES

Explanation of pic: Why different sizes?

• In this picture, we have Sample 1 and Sample 2.
During the DNA digestion step, we use restriction ● 2 - 10 BP long (Microsatellites)
enzymes to cut out sites. Let us focus on Recognition ● Made up of a unit that can vary. This unit is repeated 5 to
Site A. A represents a sequence. 100 times at each microsatellite locus
● Thousands of different STRs (short tandem repeats) are
• As you can see here, A in Sample 1 is much shorter
randomly scattered throughout the genome, but not in a
than A in Sample 2. What does that tell us? That tells
specific area
us that even if A has exactly the same sequence,
what differs is the number of times A repeats or the
number of times the sequence can be seen.
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Explanation of pic:
• TH01 represents the TCAT sequence. These are
short tandem repeats or microsatellites because it is
2-10 BP long.
• One of the most famous and common sequences
used in DNA fingerprinting is the TCAT called TH01.
• When referring to TH01, it refers to the TCAT
sequence. The number of times that it repeats is the
basis for doing DNA fingerprinting using TH01.
For TH01, it does not mean that when we do DNA
fingerprinting, it is just one sequence used when doing
DNA fingerprinting. In the forensics lab, they are actually
using more than 1 DNA sequence/ junk DNA sequence
when performing DNA fingerprinting/profiling. TH01 is just
one of the sequences used.
● Many STR sequences have only 10 to 20 alleles - thus,
cannot provide unique identification alone. (During actual
DNA fingerprinting, not only 1 DNA sequence used, many
are used)
● However, many STR loci are available and if analyzed
simultaneously, can provide enough data that the pattern
would be unique for each individual (the more DNA
sequences tested, the more specific the results and can
be enough to make sequence unique for that person
alone)
● Possible through multiplex PCR (simultaneously test on
a number of DNA sequences)
Explanation of pic:
• TH01 represents TCAT. When looking at the results
of DNA fingerprinting, each individual has a number
which corresponds to the TH01 sequence.
• Suspect A has a TH01 locus (5-3) while Suspect B
has a TH01 locus (6-10).
• What do these numbers represent and why are there
two of them?
o For Suspect A, (5-3) the TCAT was repeated five
times but also repeated 3 times.
o For Suspect B, (6-10) the TCAT was repeated 6
times and repeated 10 times.
• Why are there 2 numbers per suspect? Because
each individual inherits from both of their parents.
The 5 from one parent and the 3 from the other
parent (Suspect A). The same goes for Suspect B.
• What is its importance? For forensic cases, and is
very useful for paternity cases. Because you can
know the possible TCAT sequence of parents or the
offspring by comparing TH01 locus.
● Three different STR loci are amplified using PCR primers.
Each set of primers is labeled with a different fluorescent
label to distinguish each locus from the other. The PCR
products are run on an agarose gel to determine the
length of the fragment and therefore the number repeats.
KENNY. LOPEZ. PANOREL. PISCADERO. ROLLENAS. ROMA. SANICO. SO. SULAPAS. TAMAYO. VERGARA. VILLAROYA
Explanation of pic:
In this picture, this is the multiplex PCR in action. Three
DNA sequences are simultaneously run then it proceeds
to gel electrophoresis. This is one of the advancements in
DNA fingerprinting from restriction enzymes step in DNA
digestion step to PCR step to now doing simultaneous
testing using multiplex PCR.
● The development of PCR revolutionized DNA profiling,
making it possible for scientists to generate DNA profiles
from:
○ Trace samples - bulb of single hair strand or a
few cells from a bloodstain
■ Is important, in a crime scene not all
the time many samples can be
collected, like in a pool of blood waiting
for us, or the suspect was able to leave
behind a significantly large amount of
DNA. Being able to use the DNA in
trace samples is very useful.
○ Samples that are old and degraded - bone
found in a field or an ancient Egyptian mummy
■ In forensics, useful for cold cases
(cases not solved for a very long time)
and anthropology studies (mummies in
Egypt)
■ DNA fingerprinting can still be done
even if samples are old and degraded
● PCR-based STR is used by majority of human forensic
DNA profiling
● In an STR kit, each primer set is tagged by one of four
fluorescent dyes. Each primer set is designed to amplify
DNA fragments, the sizes of which vary depending on the
number of repeats within the region amplified.
● The sizes of the amplified DNA fragments allow scientists
to differentiate between products with labels of similar
color.
● After amplification, the DNA sample will contain a small
amount of the original template DNA and a large amount
of fluorescently labeled amplification products.
● After amplification, the DNA sample will contain a small
amount of the original template DNA sample and a large
amount of fluorescently labeled amplification products.
● Modern methods now measure the amplicons through
capillary gel electrophoresis
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○ New in here, through the advent of technology,
instead of gel electrophoresis step used to
separate fragments, capillary gel
electrophoresis is used which is much more
advanced
● Data are then analyzed by software that calculates both
the sizes of the fragments and their quantities, which are
represented as peaks on a graph
○ Instead of using Southern blot technique where
the results are still in the nylon sheet/paper, and
visualization is done in a radio radiograph, in this
one, it is analyzed directly by software where
results is shown in the computer
○ Instead of results shown as bars on a well in the
gel electrophoresis, this one will look like a
graph having peaks and troughs.
● The DNA profile can then be directly compared to a profile
from another person, from crime scene evidence, or from
other profiles stored in DNA profile databases
○ With this, it can be connected directly to the
database, for each department they might have
a database for previous suspects or any person
whom they collected DNA samples from before
○ The results of DNA fingerprinting can be
compared to their database to see if there is a
match (it’s a match!) (Usually seen on TVs)
○ Same as information systems which holds DNA
of different people whose DNA samples are
collected
○ From PCR to multiplex PCR and from gel
electrophoresis to capillary gel electrophoresis
POLYMERASE CHAIN to amplify specific DNA
REACTION fragments
CAPILLARY GEL to separate the fragments
ELECTROPHORESIS and software visualization
Because there is PCR and capillary electrophoresis, the steps
are reduced to two instead of four. In capillary electrophoresis,
there is already software and computers so the separation of
fragments and visualization of results are done as one.
KENNY. LOPEZ. PANOREL. PISCADERO. ROLLENAS. ROMA. SANICO. SO. SULAPAS. TAMAYO. VERGARA. VILLAROYA
Explanation of pic:
These are the steps. Below there is the DNA profile using
capillary electrophoresis. Instead of bars seen, peaks and
troughs of the graph are observed.
● Relative size ranges and fluorescent dye labeling colors
of 24 STR products generated by a commercially
available DNA profiling kit (shown in picture above). The
scale at the bottom of the diagram indicates DNA
fragment size in base pairs. The PCR product sizes are
revealed from the peaks of fluorescence on an
electropherogram (from Capillary Electrophoresis.
Explanation of pic:
TH01 again is one of the many sequences used by
laboratories in DNA profiling. Different sizes and colors
because of fluorescent dyes. With capillary
electrophoresis, instead of radiograph, the results
(graphs) are called an electropherogram.
4. NGS DNA Fingerprinting (Next Generation
Sequencing)
NGS Parallel sequencing of DNA into flowcells
Out of the four, this is the most advanced and newest. This is
the best in giving a DNA profile.
● NGS focuses on the actual identification of a DNA
nucleotide sequence of a gene rather than looking for
variations
○ Not merely looking for patterns or variations, but
the actual identification of a DNA nucleotide
sequence
● Able to identify down to the gene level
NOTE!
This is a sort of timeline. Again, do NOT memorize any
dates or this one. This is just to show the flow of the
advancement of technology.
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Explanation of pic:
From gel-based sequencing, to Capillary electrophoresis
which is our first-gen in the early 2000s, followed by Next-
gen sequencing then Third-gen sequencing. In this
picture, 2015 is the latest year.
● Shows the different technologies and brands of NGS
currently used
● Examples of NGS Technology used:
○ 454 Genome Sequencing System
○ Illumina MiSeq System
○ Ion Torrent System
● Steps of a sequencing experiment. Black arrows indicate
steps that are common for all second-generation
sequencing (SGS) technologies, white arrows refer to the
Illumina systems, and grey arrows refer to the Roche 454
Explanation of pic:
• Flowchart shows the different steps that each NGS
technology has. They differ in some steps and are
similar in others,but all for DNA fingerprinting using
NGS.
• NGS is very advanced as it can beat the results of 50
capillary electrophoresis. Imagine, capillary
electrophoresis is already a more advanced method
of gel electrophoresis.
• NGS is very costly as such it is not practical to use
for routine or regular forensic or paternity cases.
Impractical to use in cases that can still be solved
using STR-based DNA profiling or other DNA
fingerprinting methods.
• That is why STR is still commonly used in labs
because in NGS, since it looks at the actual
sequence instead of variations, it is more advanced
and more expensive that is why it is used for medical
research purposes such as cancer studies,
specifically those that need sensitive and specific
equipment, testing and results.
• For forensic and paternity cases, STR-based is good
enough and practical to use.
KENNY. LOPEZ. PANOREL. PISCADERO. ROLLENAS. ROMA. SANICO. SO. SULAPAS. TAMAYO. VERGARA. VILLAROYA
WHY DO WE DO DNA FINGERPRINTING?
Biological Diversity and conservation studies
disciplines: among species
Medical uses: Organ transplant compatibility testing
Hereditary disease studies
Civil Forensic analysis
investigations: Parentage and kinship testing
Verification of military casualties
Anthropological Human origins and diaspora,
studies: migration, effects of adaptation,
susceptibility and resistance to
disease
● Example of Forensic Analysis
● Comparing two suspects’ DNA fingerprints to the one
found from the crime scene. The DNA fingerprint for
Suspect 2 matches that taken from the crime scene.
Explanation of pic:
• We can't just base on the DNA fingerprinting to
directly conclude the case. It just narrows down the
possible suspects, to who was present in the crime
scene or who was in contact with the alleged victim.
• In the picture above, there are two suspects DNA
being compared and crime scene DNA. During crime
scene investigation, DNA is collected and tested and
also the DNA of the suspects, if they have.
• If there are no suspects, crime scene DNA can be
compared to their database, so probably they will hit
a match.
• Looking at the results, suspect 2 matches perfectly
found in the crime scene. Probably crime scene DNA
belongs to Suspect 2, while Suspect 1 is ruled out
because it is very different.
• Since it is a laboratory test, which one is known and
which one is unknown? Which one are we doing
testing on? Unknown samples would be crime scene
DNA because we don’t know whose DNA it belongs
to while suspects’ DNA are known samples.
• If it comes from a database, you already know that
the DNA sequence belongs to this person. If you are
doing actual testing, you will know that the DNA
belongs to Suspect 2.
• You are trying to see if your unknown matches one
of the knowns just like what we do in our different
laboratory tests. Comparing the unknown to the
knowns that we have.
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• Thus, a forensically useful material should be:
o Unique
o Not change over time (i.e., during normal use)
o Likely to be left at a scene in sufficient quantity to
establish a match
o Not change after being left at the scene and during
subsequent examination
• As an investigator, PPE should be worn to avoid
contamination in the crime scene with your own DNA.
• Of all evidence left in the crime scene, DNA fingerprints
are the most important. Of these requirements, only DNA
fingerprints qualify– especially with the advent of PCR,
● Example of Paternity Testing even trace minute samples of DNA can be amplified to
● The DNA profiles of two parents are compared with their produce DNA fingerprints that can be then visualized
child’s. The STRs in the child have been inherited from through capillary electrophoresis.
which parent is apparent. • Thus, DNA profiling is considered the gold standard of
Forensic Analysis. When we talk about forensic analysis,
Explanation of pic:
it does not automatically mean DNA fingerprinting. There
• TH01 which repeats TCAT. How do they determine
are still other tests or methods that are done in forensic
paternity in a paternity case? Rule out2 gihapon.
analysis.
• Let us assume this is a very simplified
electropherogram. Let us assume this is TH01 so
TOPIC OUTLINE
TCAT sequence the child has 5 and 7. Again, all of
us have 2. WHAT is DNA A form of individual identification at the
• The 5 in the child matches the 5 in the DNA sample fingerprinting? DNA level using molecular means
of the mother because the mother has 5-10. Father
has 7-13 and the 7 of the father matches to the 7 of WHAT do we do? Look at the variation of sequences and
that variation makes each pattern
the child. We can assume here that this child is really
unique enough to establish an
the offspring of the father and mother. identification
• The only possible offspring that the father can
produce, and the mother can be seen on the child. WHO discovered Dr. Alec Jeffreys in 1985
• Then of course, if they have other children, 10-13, 7- it?
10 or any other different combinations as long as 5-
10 is from mother’s side while 7-10 from father’s side. HOW is it done? RFLP based DNA fingerprinting
VNTR based DNA fingerprinting
• Can a person inherit the same number of repeated STR DNA Profiling
sequences from both parents? Yes, for example, the Next Generation Sequencing DNA
mother is 5-10 and the father is 7-10, it is possible Profiling
that both 10s can be inherited by the child having a
TH01 locus of 10-10. That is not an anomaly, but it is WHY do we do Forensic Analysis
a possible result if there is a number that is common it? Organ Transplant Compatibility Testing
Human Origin and migration studies
between both parents.
Paternity Testing

For WHOM is it? Medical personnel

WHY IS DNA FINGERPRINTING ESSENTIAL IN FORENSIC (The people who Researchers
ANALYSIS? are working in Investigators
Most forensic disciplines concerned with offenses against the these fields) Environmentalists
person, and some other crimes, try to establish a link between
items found at the scene and items found on or associated RFLP and VNTR use VNTRs minisatellites while STR uses
with a suspect. In other words, to establish whether the STRs or microsatellites. The latest NGs which is very
recovered items could have originated from the same source advanced and expensive are not really used in forensics but
(through DNA fingerprinting). This process can be more on research and medical studies.
summarized as:
● Establishing a match Make sure to keep this lecture in mind because it will be useful
● Calculating the significance of the match in the third year :)

NOTE! Unsay tawag sa kuto ng kalbo?

Again, we are just trying to establish a link and not a final Ano?
answer or a conviction. Homeless. WAKAKAKAAHAHAHA

• The goals of forensic matching are to reduce the potential

population from which an item could have come to one
individual within the population.

KENNY. LOPEZ. PANOREL. PISCADERO. ROLLENAS. ROMA. SANICO. SO. SULAPAS. TAMAYO. VERGARA. VILLAROYA 9