FORENSIC DNA

ANALYSIS

Ms. Joanne S. Llacer

FORENSIC DNA ANALYSIS

DNA Overview
Sources of Biological Evidence
Documentation & Labeling
Collection and Preservation
Steps in DNA Analysis
Limitations of DNA Analysis

OUTLINE
 LEARNING OBJECTIVES

Upon successful completion of subject,

the student should be able to do the
following:
•Define what is DNA and appreciate the
importance of its analysis in crime
investigation;
•Explain the importance of maintaining the
integrity of physical evidence.
•Describe methods used to locate evidence at a
crime scene.
 LEARNING OBJECTIVES
• Describe the importance of scene documentation
in successful case resolution.
•Enumerate priorities for collecting biological
evidence.
•Define situations that require control and
reference samples to be collected.
•Describe equipment used when collecting
biological evidence.
•Describe procedures used to collect, mark and
package wet and dry biological evidence
DEOXYRIBONUCLEIC ACID
•DNA is your genetic blueprint.
•You inherit it from your biological parents.
•It codes for things like eye color, hair color,
stature, predisposition to some diseases and
many other human traits

SCHEMATIC REPRESENTATION
 INDIVIDUAL
46 CHROMOSOMES
23 pairs (22 pairs or chromosomes + 1 pair sex
chromosomes)

MOTHER FATHER

46 Chromosomes for Normal Human Beings

 Normal Male

Shows 23 pairs of chromosomes. Twenty two pairs are called autosomes, 1 pair is
called the sex chromosomes ; XY in a male. Males are associated with secondary
sexual characteristics – abundant facial hair, voice, others. Differences are also
evident in the genitalia.
 Normal Female

Shows 23 pairs as in the male, but sex chromosomes are represented by XX. Females also
have different physical features from males. A Barr body is also present in cells of the
female, representing an inactivated X chromosome.
Sources of Biological Samples:
 DNA OVERVIEW
FROM THE WHOLE TO THE MICROSCOPIC PARTS
Diagram of a Typical Cell from Human

Double-stranded DNA
is organized into
chromosomes.

Chromosomes are
situated in the nucleus
and the membrane
bound nucleus is found
in the cell.
DNA in the Cell

Target Region for PCR

 Numbers and DNA
Number of cells in human body 100 trillion
Number of Chromosomes 23 pairs
Length of DNA in each cell 2 meters
Size of genome in base pairs 3 billion
Length of DNA in a person 150 trillion m
Distance from earth to sun 150 billion m
Each person has enough DNA to go to the sun and
back 500 times!!
 STEPS IN DNA ANALYSIS

1. Collection of sample 5. Running of samples for

Visualization

2. Documentation &
Screening 6. Data Interpretation and
Report Making
3. Extraction of DNA
7. Releasing of Report
4. DNA Amplification
Brief History of Forensic DNA Typing

▪ 1988 - FBI starts DNA casework

▪ 1995 - FSS starts UK DNA
database
▪ 1998 - FBI launches CODIS
database
▪ 2001 - PNP opens DNA Lab
▪ 2007 – Rules on DNA Evidence
approved by RP Supreme Court
 Applications of DNA Analysis

• Criminal Investigations -
matching suspect with
evidence
• Parentage Testing - identifying
parent(s)
• Historical investigations
• Missing persons investigations
• Mass disasters – positive
identification of the deceased
• DNA databases
DNA ANALYSIS IN CRIME INVESTIGATION

• Each person’s DNA is

different from every
individual’s, except for
identical twins.
• DNA collected from a
crime scene can either
link a suspect to the
evidence or eliminate a
suspect, similar to the
use of fingerprints.
 How do DNA evidence work?
In this example, the pattern of the evidence
specimen matches that of suspect number one.
 DNA in Criminal Investigations

•Skin, blood, semen, teeth, muscle, fingernails,

bone, feces, hairs and saliva all contain DNA.
•The same DNA is found in all cells of the body.
•The same DNA profile is recovered, no matter
what the sample (cell) type.
•Your DNA does not change through your life.
•Any cellular material left at a crime scene may
be a useful source of DNA.
 HUMAN IDENTITY TESTING
•Criminal Investigations – matching suspects(s)
with evidence
•Convicted offender DNA databases
•Mass disasters – positive identification of the
deceased
•Missing persons investigations
•Parentage testing – identifying parents(s)
•Historical investigations
•Military DNA “dog tag.”
 SEARCH FOR DNA EVIDENCE

▪ DNA Evidence is Transfer

Evidence.

▪ DNA Evidence is Trace Evidence.

 DNA Evidence is Transfer Evidence
Evidence Possible Location of Sources of DNA
DNA evidence
Baseball bat or Handle, end Sweat, skin, blood
▪ similar weapon
Hat, bandanna, Inside Sweat, hair, dandruff
mask
Facial tissue, Surface area Mucus, blood, sweat,
cotton swab semen, ear wax
Dirty laundry Surface area Blood, sweat, semen
toothpick Surface area Saliva, blood
Fingernail, partial scrapings Blood, sweat, tissue
fingernail
Through and Outside surface Blood, tissue
through bullet
eyeglasses Nose or ear pieces, lens Sweat, skin
 DNA Evidence is Transfer Evidence
Evidence Possible Location of Sources of DNA
DNA evidence
Used cigarette Cigarette butt Saliva
▪
Stamp or envelop Licked area Saliva

Tape or ligature Inside /outside surface Skin, sweat

Bottle, can, or Side, mouthpiece Saliva, sweat,
glasses
Bite mark Person’s skin or clothing saliva
Used condom Inside/outside surface Semen, vaginal or rectal
cells
Blanket, pillow, Surface area Sweat, hair, semen, urine,
sheet saliva
G:\DNA\LivePicture\S\K0060c_p_K0177.jpg

tissue fingernails

cigarette butt Broken window

 DNA Evidence is Trace
Evidence
Evidence such as hair, fiber,
body fluids, are types of
physical evidence that is
small and transient, but
measurable. When larger
items of physical evidence
are subjected to closer
examination in the
laboratory, trace evidence
may be detected.
DNA Evidence Collection
And
Preservation
 Collection Priority

▪ The first Priority are trace materials

and evidence of a fragile nature.
▪ The second priority would be to
collect swabs from handled items
that have been moved, are out of
place or do not belong to the
resident.
▪ A third priority type of evidence
that may be at the scene includes
the potentially lower-quality
biological evidence.
 COLLECTION OF DNA
EVIDENCE
“As a general rule – air
dry any sample taken.
If it is in the form of
tissues, either freeze or
soak in lysis buffer
solution.”
▪ “If DNA evidence is not
properly documented,
collected, packaged, and
preserved, it will not meet
the legal and scientific
requirements for
admissibility in a court of
law”.
- If DNA evidence is not
properly documented, its
origin can be questioned.
- If it is not properly
collected, degradation
can occur.
- If it is not properly
packaged, contamination
can occur.
- If it is not properly
preserved, degradation
can occur.
 GENERAL
GUIDELINES
Degradation of biological evidence is
most commonly caused by exposure to
moisture and heat, which encourages the
growth of bacteria.

This can lead to destruction of the

DNA and the possibility of incomplete or
no DNA typing results being obtained.
 CONTAMINATION OF DNA EVIDENCE

Contamination of
samples, either by
coming into contact with
each other, or with DNA
from a person handling
the sample, can lead to
un‐interpretable DNA
results.
 DNA Source Example: Ashtray

▪ In this example, DNA

evidence left by the
suspect and/or the victim
could be found in the
form of saliva, skin cells
and sweat on the
cigarette.
 DNA Source Example: Laundry

In this example, DNA

evidence from both the
victim and suspect can be
found in the form of sweat,
saliva, blood and/or semen.
 DNA Source Example: Baseball Bat
▪ At first glance, the victim's
DNA evidence (hair, blood
and bone fragments)
would be found at the tip
of the baseball bat. After
further consideration, DNA
evidence left behind by the
suspect (sweat and skin
cells) could be found on
the handle of the bat.
 Non-traditional Locations of
DNA Evidence

▪ A threatening letter was sent to a

newspaper editor. The FBI Lab
sampled the envelope flap and
recovered some cells (saliva)
that were then typed for DNA.
The profile from the envelope
flap was compared to a known
suspect’s profile and was found
to match.
Non-traditional Location Example: Saliva
▪ In 1997, two women from Florida were victims of sexual
assault and robbery. One year later, the police developed
a suspect. Plain-clothed police officers monitored the
suspect for months looking for clues that would build
their case. During surveillance, the officers saw the
suspect spit on the street. One of the officers grabbed a
napkin and collected the spittle. The saliva, which
contained cells, provided enough DNA evidence to
charge the man with the two attacks.
Non-traditional Location Example: Gunshot
A suspect in the violent robbery of a shop owner.
While the owner of the shop shot at the suspect in self-
defense, the bullet from his gun went through the body
of the assailant and lodged into a wall. When the
ballistics team evaluated the projectile, they discovered
traces of blood on its surface. Upon DNA analysis, the
blood on the projectile recovered from the crime scene
was proved to be that of the suspect, who previously
claimed he knew nothing of the incident.
 Personal Protection
BIOLOGICAL MATERIALS CAN CONTAIN PATHOGENS SUCH
AS:
•Hepatitis
•Syphilis
•TB
•Gonorrhea
•Measles
•HIV

NOTE: Assume that all stains, wet or dry, are infectious!

 Considerations for Handling
1. Always wear latex gloves when handling biological
evidence.
2. Change gloves between handling of each item of
evidence.
3. Process biological evidence on a clean area (such as a
fresh piece of butcher paper). Use a 10 % bleach
solution to clean areas at which biological evidence
will be handled.
4. Be careful not to talk excessively over biological
evidence. Wear a mask if necessary.
5. Only work with one piece of evidence at a time.
6. Never allow items of biological evidence to come into
contact with each other.
 Buccal Swab
Sterile swabs or other buccal collection devices
are rubbed against the inside cheek of the
individual's mouth to collect cells for analysis.
Procedure:
• Collect oral swabs from possible suspect(s),
victim(s), and known references (those with
access to the scene, such as homeowners).
• Do not pre-wet swab.
• Rub dry swab on the inside of cheek until wet.
Collect at least two swabs from cheeks.
• Identify item with donor's name on swab box
and envelope.
• Thoroughly air-dry the swab before packaging.
 Liquid Blood Sample Procedure
• Collect in purple-topped vacuum
tubes that contain the preservative
ethylenediamine tetra acetic acid.
• Identify item with donor's name on
blood tube.
• Refrigerate. Do not freeze or store
near high heat (above 100° F) to
prevent the glass from fracturing.
• Alert evidence officers of
refrigerated samples.
 Cigarette Butts
▪ Select only the cigarette
butts that may be of
evidentiary value
▪ Insure that the cigarette
butts are thoroughly air
dried before packaging.
▪ Do NOT handle with your
bare fingers.
▪ Do NOT include the ashes.
Blood and Body Fluid
Collection
▪ Cuttings.
Remove a
section of the
item containing
the stain using a
sterile or clean
cutting device.
Blood and Body Fluid Collection
Wet absorption.
• Moisten sterile swab
with distilled water.
• Press or rub the swab
into the stain.
• The stain should be
concentrated on the tip.
• Allowed swabbed stain
to air-dry.
 Blood and Body Fluid Collection
▪ Scraping method. The sample is scraped with a clean razor blade or
scalpel, into a clean piece of paper that can be folded and packaged
in a paper envelope. This is a method to be used in a controlled
environment (i.e., no wind or traffic) and where the scrapings will not
contaminate other evidence.
Hair

• Collect hair using

clean forceps.
• Place hair
evidence into a
clean piece of
paper that can be
folded.
• Package in a
paper envelope.
Wet Stains on Absorbent
Surfaces
▪ Use one or more dry sterile cotton swabs to soak
up the stain.
▪ If the stain is small, concentrate the stain on the
tip of the swab.
▪ Thoroughly air-dry the sample.
▪ Document that the stain was found wet and
identify from where the stain
▪ Submit entire item if swabbing is not possible.
Special Situations

Procedures for collecting

wet stains on absorbent
substances are as follows:
1. Scoop a thin layer of
soil (or sand).
2. Allow to thoroughly
air-dry.
 Special Situations
For Liquid Containers:
▪ Empty liquid containers by poking a hole in the
bottom to avoid liquid contact with the mouth
area.
▪ Swab mouth area of the container using sterile
cotton swabs. Package in paper envelope.
▪ Submit entire item if swabbing is not possible.
▪ Request test for both DNA and latent fingerprints.
▪ Mark aluminum cans for "Room Temperature
Storage," as cold or frozen storage causes
condensation on metal, which may dilute biological
evidence.
Wet Stains on Nonabsorbent
Surfaces
▪ Use one or more sterile swabs (or sterile gauze
for larger stains) to soak up the stain.
▪ Concentrate the stain on one portion of the tip of
the swab.
▪ Allow to thoroughly air-dry.
▪ Collect the control sample by slightly moistening
the swab with distilled water and rubbing an area
of the surface in an unstained region near the
stain.
 Dry Stains on Absorbent
Surfaces
▪ (examples: wood frame and carpet)
▪ Cut out the stained area and package in paper.
▪ Collect a portion of the unstained area as a
control sample.
▪ Package and label each sample separately.
 Dry Stains on Nonabsorbent
Surfaces
▪ Swabbing method (wet absorption).
 Moisten sterile swab with distilled water.
 Concentrate stain on the tip of the cotton swab.
 Allowed swabbed stain to air-dry.
▪ Scraping method.
 Use a new or clean scalpel blade to scrape the stains from the surface.
 Collect the flakes onto clean paper and fold the paper in a bindle.
 If the stain is on wood, shave the area of the bloodstain with a new or clean
scalpel blade.
▪ Package each item separately.
▪ Place sample in a labeled envelope that provides reference
information on where the sample was collected.
▪ Take a control swabbing from unstained areas using a new sterile
swab slightly moistened with distilled water. Allow the control
swab to air-dry, label and package in paper.
▪ Submit entire item if swabbing or scraping is not possible.
• Collect samples from the victim
to act as control
 Documentation & Labeling

DOCUMENTATION – Refers to any written or printed matter conveying

authoritative information, records, reports or evidence.

IMPORTANCE:

a. To maintain proper chain of custody (this refers to the markings,

labels, and records made on the specimen from the crime scene to the forensic
laboratory).

b. To preserve the original condition and position of evidence at the

crime scene. (e.g. photograph, sketch)

*** If DNA evidence is not properly documented prior to collection, its

origin can be questioned thereby it will NOT meet the legal and scientific
requirements for admissibility into a court of law.
 Documentation & Labeling

DOCUMENTS NEEDED

1. LETTER REQUEST
2. COURT ORDER
3. CHAIN OF CUSTODY FORM (Ideal)
 Documentation & Labeling

CHAIN OF CUSTODY

record of individuals who have had physical

possession of the evidence.

Note: Integrity of chain of custody

The fewer people handling the evidence, the better, the

lesser chance of contamination and a shorter chain of
custody for court admissibility hearings.
 Documentation & Labeling
GUIDELINES FOR DOCUMENTATION OF DNA EVIDENCE
1. Evidence at the Crime Scene:
a. Photograph and/or videotape the evidence before touching, moving or collecting
it.

b. Record the location and condition of the evidence. (Humidity, exposure to sunlight,
chemicals, etc.)

c. Note and sketch the evidence in relation to the crime scene and other object
present.

d. Record the condition of the biological evidence during the time of collection.
 Documentation & Labeling

2. Markings on Evidence during the collection

Markings on the specimen must at least

contain the following:
A. Exhibits

B. Initials and or signature of the

collecting officer.

c. Time and date of collection.

 Documentation & Labeling

a. Exhibits

b. Initials and or signature c. Time and date of

of the collecting officer. collection.
 Documentation & Labeling

3. Place or site of markings:

a. STAINED CLOTH, GARMENTS, SEAT
COVER, ETC.
Markings on the specimen must be
placed far from the portion of interest
such as blood or seminal stains so as not
to contaminate the samples. Markings
on the paper bag used to contain the
specimen must also be taken into
account.
Documentation & Labeling
Documentation & Labeling
Documentation & Labeling

b. BLOOD SAMPLES
It must be placed in a clean airtight container or test
tubes where the markings can be made.
 Documentation & Labeling

c. HAIRS, FINGERNAIL
CUTTINGS, BLOOD
SCRAPPINGS, COTTON
SWAB, ETC.
Markings must be made on
the container or enveloped
that was used to contain the
specimen. To include the
description and the number
of specimen contained.
Documentation & Labeling
 Documentation & Labeling

Labeling through Tagging

Tagging could be applied to movable objects such as this

knife - where marking/s placed on any of its blade surface
is not possible.
Documentation & Labeling

Location of
recovery

The tag must contain the markings and other

information about the specimen.
Documentation & Labeling

It is advisable to place same

markings such as exhibit and
initials on a secure portion of
the specimen for the
following reasons:

a. possibility for the tag to be

dislodged or removed during
loose tagging or incautious
transport of the specimen;

b. to further intensify
identification of the specimen if
ever the need arises (e.g. court of
law/proceedings)
 Documentation & Labeling

d. TISSUE SAMPLES (SKIN, EAR

TISSUE, MUSCLE TISSUE,
COAGULATED BLOOD, ETC.)
Markings are to be placed on
a clean airtight plastic or glass
container used.
Documentation & Labeling
 Documentation & Labeling

*** It is advised that permanent marker or ink must be

used.

*** It is required that rubber gloves must be worn at

all times during the conduct of markings (BSI, CONT).
Documentation & Labeling

GUIDELINES FOR LETTER

REQUEST
Ideal request sample containing all
information as to:

-brief summary of the case

-source of the specimen

-markings

-packaging
 Collection and Preservation
Personal protection:
BIOLOGICAL MATERIALS CAN
CONTAIN PATHOGENS SUCH AS:
•Hepatitis
•Syphilis
•TB
•Gonorrhea
•Measles
•HIV
NOTE: Assume that all stains, wet or dry, are
infectious!
GUIDELINES for COLLECTING
SAMPLES for DNA Examination

➢ Must be properly documented

➢ Must be properly collected
➢ Prevent contamination
➢ Avoid enzymatic/microbial attack
➢ Avoid excessive sun exposure
➢ Collect and keep samples/specimen at least in
duplicate
➢ Collect samples from the victim to act as
control
Has Forensic DNA Profiling
replaced other forensic techniques?

•Not all cases can be solved with DNA.

•Not all cases can be solved with
fingerprinting.
•The disciplines can work in synergy.
•Forensic science is multidisciplinary:
pathology, biology, anthropology,
odontology, toxicology, ballistics,
fingerprints, etc.
DNA cannot be used to:

•Tell when the suspect was at the crime

scene.
•Tell how long the suspect was at the
crime scene.
•Tell how long the suspect handled an
object.
•Determine how long ago the cells were
deposited on the item.
How long does crime scene
DNA last?
•If items have been stored properly, DNA
can last decades (30+ years).
•Exposed to harsh environment (eg: heat,
sunlight, moisture, mold), DNA may only
last a few days or weeks.
•In the laboratory: we can’t tell in advance
whether or not an item will yield any DNA.
•DNA can be present, but too degraded to
give a result (partial DNA profiles)
Thank you!!!