CHAPTER 14

DEOXYRIBONUCLEIC ACID (DNA)

(DEOXYRIBONUCLEIC ACID) 

During the mid 1980's DNA analysis was first recognized as having application to
forensic science by the British molecular biologist Alex Jeffreys. From work in his laboratory, as
well from others, it was realized that DNA has been utilized as a new powerful tool for human
identification. It offers the following advantages: 
1. DNA is stable – it can be isolated from material that is months or even years old. 
2. DNA can be destroyed from wide variety of biological resources like blood, 
semen, hair, saliva and bone.
3. DNA can be replicated in the laboratory – from a very small amount of initial 
material through the process of PCR (POLYMERASE CHAIN REACTION).
4. DNA shows greater variability from one individual to the next. 

WHAT IS DNA? 

DNA is functionally the hereditary material that contains the genetic information
necessary for the duplication of cells and for the production of proteins. Chemically, it is an acid,
is phosphorous rich, contains a deoxyribose sugar, and contains the four bases which show a
unique property of pairwise equivalency. It is a double helix composed of two complimentary
strands. 

FACTS ABOUT DNA

1. DNA - de-oxy-ribo-nu-cle-ic-acid 
is a chemical substance found in all cells whose composition have been passed on
from parents to their children through DNA replication. All cells in the body
have the same DNA composition except individual egg and sperm cells. 

In DNA replication, the two strands of the DNA double helix unwind, the separated strands serving as templates for the
formation of new DNA strands. Free nucleotides
pair with the complementary bases on the separated strands of DNA. This process ultimately results in the complete replication of
the DNA molecule.

2. BIOLOGICAL EVIDENCES that can be submitted for DNA analysis: 

a. Blood and bloodstain

b. Semen and seminal stain
c. Hairs and follicles or root
d. Saliva or buccal swab
e. Bones and organs
f. Tissues and cells 
3. LINE-UP OF CASES where DNA ANALYSIS can be of help: 
a. Sexual assault case like rape
b. Murder
c. Homicide
d. Robbery
e. Hit and run
f. Extortion
g. Paternity case
h. Identification of remains from mass disaster cases and missing persons 

4. HOW DNA ANALYSIS is used to identify with accuracy the perpetrators of crime: 

Human tissues such as hair, blood, semen is often left in places where a crime has
been committed. By carefully collecting such bits of tissues, their owner can be identified
from the DNA pattern obtained. 

GUIDELINES FOR THE COLLECTION AND PRESERVATION OF SAMPLE

EVIDENCE FOR DNA ANALYSIS 

There are hundreds of varieties of physical evidence commonly submitted for examination for
forensic science laboratories by law enforcement agencies. Evidence that could be subjected to
DNA analysis is generally limited to substance that are been successfully isolated and analyzed: 

1. Blood and bloodstains 4. Hairs and follicles  

2. Semen and seminal stains 5. Tissues and cells
3. Saliva and buccal swab  6. Bones and organs 

In addition, there are reports indicating that DNA has been isolated from urine samples
with nucleated cells; however, it is extremely rare to be able to obtain sufficient DNA type from
urine samples. Other types of biological evidence, such tears, perspiration, serum and other body
fluids without nucleated cells are not amenable to DNA analysis. It should be kept in mind that
not all biological materials listed above in case work submitted to a forensic laboratory are in
such a state that DNA can be successfully extracted and analyzed. 

When collecting any type of body fluid or tissue, the universal precaution for the body
fluids should be taken. Wear gloves anytime, these specimens are handled with additional
protective equipment when appropriate. All body fluids and tissues must be assumed to be
infectious regardless of the source. 

COLLECTION AND PRESERVATION OF BIOLOGICAL EVIDENCE 

The ability to perform successful DNA analysis on biological evidence recovered from
the crime scene depends very much on what kinds of specimens were collected and how they
were preserved. Thus, the technique used to collect and document such evidence, the quantity
and type of evidence that should be packaged, and how the evidence should be preserved, are
some of the critical points for a forensic DNA testing program. Unless the evidence is properly
documented, collected, packaged and preserved, it will not meet the legal and scientific
requirements for admissibility into a court of law. If the DNA evidence is not properly
documented prior to collection, its origin can be questioned. If it is improperly packaged, cross
examination may occur. And if the DNA evidence is not properly preserved, decomposition and
deterioration may well occur. Any of these effects will seriously affect the outcome of DNA
typing. The following are general guidelines for the documentation, collection and packaging
and preservation of DNA evidence. 

GUIDELINES FOR DOCUMENTATION OF DNA EVIDENCE 

The initial stages in physical evidence examination encompass activities that take place at a
crime scene as well as the forensic laboratory. Documentation is important from two points of
view in forensic science; the legal one, and the scientific one. Nothing should ever be altered
until its original condition and positions have been recorded. Several different means of
documentation are available. Generally, the use of more than one method is recommended.
Every major piece of evidence should be documented. 

COLLECTION  OF DNA EVIDENCE AT THE CRIME SCENE 

EVIDENCE CONDITION  LOCATION COLLECTION

Blood  Liquid  Person Collect in EDTA tubes

Use syringe, collect
in EDTA tubes.
Liquid  Scene 
Transfer onto cotton cloth.
Air dry
Collect cloth in test tube.
Blood  Cloth  Scene  Transfer onto cotton 
cloth. Air dry.
Air dry at room
Blood  Wet Clothing  temperature. Package
in paper bag.
Air dry at room
Object 
Wet temperature. Package
in paper bag.
Collect sample with
syringe. Place sample in
Wet Water 
plastic container.
Freeze sample.
Scrape crust into paper
Blood packet. Collect
Dried Blood Crust Person, Scene, Object 
control blank.

Collect item directly.

Stain Weapon 
Collect entire item
Stain Small Object 
Cut out stained area.
Vehicle, upholstery, carpet,
Stain Package separately. Collect
wallpaper, wood 
control.
Stain Unmovable surface, concrete a.)Scrape into paper
 packet. Collect control.
wall  b.)Transfer onto
wall
moistened cotton
thread. Air dry thread.
Unmovable surface  Tape lifting. Place in
Spatters 
container.
a.)Collect sample with rape
Semen  Victim  kit. b.)Collect sample with
Liquid 
swabs. Keep in
refrigerator. 
Object, Scene  a.)Collect liquid semen
Liquid  into tube b.)Transfer onto
cotton cloth. Air dry. 
Wet  Clothing Air dry. Package
separately. 
Collect as is. Package
Dried Stain  Clothing, carpet 
separately.
Cut out section with stain.
Upholstery 
Collect control.
Scrape sample into paper
Unmovable surface 
packet. Collect control.
Direct deposit in container.
Urine  Liquid  Person 
Keep refrigerated. 

COLLECTION OF DNA EVIDENCE AT THE CRIME SCENE

 
EVIDENCE CONDITION LOCATION COLLECTION
Use syringe transfer into test
Saliva  Liquid  Scene 
tube. Keep refrigerated.
Clothing, Object  Collect as is.

Tissue, Organ  Fresh  Scene  Place in container. Keep cold.

Dried  Scene  Place in container.

Collect hair with tissue in
Bone  With Tissue  Scene 
container. Keep refrigerated.
Separate hair from blood.
Hair  With Blood  Scene 
Collect in paper packet.
Pick up sample with clean
Intact Hair  Scene 
forceps. Place in paper packet.
Fragments  Scene  Tape lift. Package in container.

Control  Person  Pulled (at least 20) 

DNA ANALYSIS 
 There are many types of DNA testing that are presently available. One detects presence
of RFLP's (Restriction Fragment Length Polymorphism) in the DNA. This is commonly known
as “DNA Profiling” of “DNA Fingerprinting" and in most cases results in either a positive of
exclusion of an individual as a donor. This analysis requires approximately 100 nanograms of
high quality DNA for a successful determination. DNA analysis in forensic casework was first
performed using this technique. In this approach, purified DNA is first cut with certain
restriction endonucleases and then run on an agarose gel. The separated DNA fragments are
subsequently blotted on to a membrane and exposed to radioactivity - labeled probes specific for
regions located between the restriction sites, which vary in length within the population.
Autobiography then reveals labeled restriction fragments,  the bonding pattern of which is used
for comparison between victim and suspect for  comparison with a database. 

The advent of PCR technology and its application to forensic science, brought a new way
of examining biological evidence and has paved the way for the other technique – the PCR
amplification and typing of the HLA DQA01 and 5 Polymarkers (PM) loci which requires only 2
nanograms of DNA. PCR analysis of biological evidence was first used in a criminal case in the
United States in 1986 and has been used in a large number of court cases and has proved a
reliable and widely accepted method for the examination of human identity. 

One of the most important developments in the field of human identity testing is the use
of DNA typing to analyze biological evidence. In particular, the powerful PCR is used to analyze
samples which cannot be typed by other methods, such as samples containing minute amounts of
human DNA and very old and/or degraded DNA. 

HOW DNA ANALYSIS IS DONE 

DNA TYPING is done by first carefully extracting the DNA from the evidentiary
samples. The DNA is then analyzed to give a particular pattern. The patterns are compared with
that of a known individual to determine a match. In individual identification, the pattern obtained
from the evidentiary sample is compared with that of a suspect. If the patterns are different the
evidentiary sample definitely has not originated from the suspect. The DNA pattern of the
evidentiary sample is similar to that of sample obtained from the suspect, the probability that the
evidentiary sample is similar to that sample arose from the suspect, and not from a random
individual in population is calculated from a formula based on well-accepted concepts of
statistical probabilities and population genetics using an established population genetic database. 
Probability calculations must show that no other person in the country or in the world
could possess such DNA pattern except the suspect. For example, the probability of a matched
DNA pattern being present in the Philippines indicates how many people are expected to possess
such pattern. If probability of pattern is 1 per 20,000, this means that there could be as many
3,600 (72 million/20,000) people having that pattern. Therefore, the DNA test is inconclusive.
However, if DNA pattern has a probability of 1 to 100 million, since there are only almost 80
million people in the Philippines, then the forensic sample must have come from the suspect.