GRADUATE SCHOOL OF CRIMINAL JUSTICE AND PUBLIC SAFETY

General Luna Road, Baguio City

Name : Jan Michael A. Fernandez

Facilitator : Gerardo K. Tumbaga, SR., Ph.D.
Subject : Advanced Techniques for Crime Analysis and
Investigation
Topic : Forensic DNA Typing
Schedule : 2:00 pm- 5:00 pm (S-Sun)
Date : September 25, 2021

Objectives

 Understand the importance of the DNA

 Identify the properties of DNA
 Understand the importance of Collection and preservation of DNA
 Understand the Mitochondria DNA
 Understand the admissibility of DNA Evidence.

Introduction

DNA, or deoxyribonucleic acid, is the hereditary material in humans and almost all other organisms.
Nearly every cell in a person’s body has the same DNA. Most DNA is located in the cell nucleus
(where it is called nuclear DNA), but a small amount of DNA can also be found in the mitochondria
(where it is called mitochondrial DNA or mtDNA). Mitochondria are structures within cells that convert
the energy from food into a form that cells can use.
The information in DNA is stored as a code made up of four chemical bases: adenine (A), guanine
(G), cytosine (C), and thymine (T). Human DNA consists of about 3 billion bases, and more than 99
percent of those bases are the same in all people. The order, or sequence, of these bases determines
the information available for building and maintaining an organism, similar to the way in which letters
of the alphabet appear in a certain order to form words and sentences.
DNA bases pair up with each other, A with T and C with G, to form units called base pairs. Each base
is also attached to a sugar molecule and a phosphate molecule. Together, a base, sugar, and
phosphate are called a nucleotide. Nucleotides are arranged in two long strands that form a spiral
called a double helix. The structure of the double helix is somewhat like a ladder, with the base pairs
forming the ladder’s rungs and the sugar and phosphate molecules forming the vertical sidepieces of
the ladder.
An important property of DNA is that it can replicate, or make copies of itself. Each strand of DNA in
the double helix can serve as a pattern for duplicating the sequence of bases. This is critical when
cells divide because each new cell needs to have an exact copy of the DNA present in the old cell.
DNA is a double helix formed by base pairs attached to a sugar-phosphate backbone.
DNA is the chemical name for the molecule that carries genetic instructions in all living things. The
DNA molecule consists of two strands that wind around one another to form a shape known as a
double helix. Each strand has a backbone made of alternating sugar (deoxyribose) and phosphate
groups. Attached to each sugar is one of four bases--adenine (A), cytosine (C), guanine (G), and
thymine (T). The two strands are held together by bonds between the bases; adenine bonds with
thymine, and cytosine bonds with guanine. The sequence of the bases along the backbones serves
as instructions for assembling protein and RNA molecules.
Body of the Report

What is DNA

DNA is the chemical name for the molecule that carries genetic instructions in all living things. The
DNA molecule consists of two strands that wind around one another to form a shape known as a
double helix.

DNA, or deoxyribonucleic acid, is the hereditary material in humans and almost all other organisms.
Nearly every cell in a person's body has the same DNA. ... The information in DNA is stored as a
code made up of four chemical bases: adenine (A), guanine (G), cytosine (C), and thymine (T).

DNA, or deoxyribonucleic acid, is the fundamental building block for an individual's entire genetic
makeup. It is a component of virtually every cell in the human body. Further, a person's DNA is the
same in every cell. For example, the DNA in a man's blood is the same as the DNA in his skin cells,
semen, and saliva.

DNA is a powerful tool because each person's DNA is different from every other individual's, except
for identical twins. Because of that difference, DNA collected from a crime scene can either link a
suspect to the evidence or eliminate a suspect, similar to the use of fingerprints. It also can identify a
victim through DNA from relatives, even when no body can be found. And when evidence from one
crime scene is compared with evidence from another, those crime scenes can be linked to the same
perpetrator locally, statewide, and across the Nation.

Forensically valuable DNA can be found on evidence that is decades old. However, several factors
can affect the DNA left at a crime scene, including environmental factors (e.g., heat, sunlight,
moisture, bacteria, and mold). Therefore, not all DNA evidence will result in a usable DNA profile.
Further, just like fingerprints, DNA testing cannot tell officers when the suspect was at the crime scene
or for how long.

What does DNA do?

DNA contains the instructions needed for an organism to develop, survive and reproduce. To carry
out these functions, DNA sequences must be converted into messages that can be used to produce
proteins, which are the complex molecules that do most of the work in our bodies.

Where Is DNA Contained in the Human Body?

DNA is contained in blood, semen, skin cells, tissue, organs, muscle, brain cells, bone, teeth, hair,
saliva, mucus, perspiration, fingernails, urine, feces, etc.

Where can DNA evidence be found at a crime scene?

DNA evidence can be collected from virtually anywhere. DNA has helped solve many cases when
imaginative investigators collected evidence from nontraditional sources (see "Identifying DNA
Evidence"). One murder was solved when the suspect's DNA, taken from saliva in a dental
impression mold, matched the DNA swabbed from a bite mark on the victim. A masked rapist was
convicted of forced oral copulation when his victim's DNA matched DNA swabbed from the suspect's
penis 6 hours after the offense. Numerous cases have been solved by DNA analysis of saliva on
cigarette butts, postage stamps, and the area around the mouth opening on ski masks. DNA analysis
of a single hair (without the root) found deep in the victim's throat provided a critical piece of evidence
used in a capital murder conviction.

Identifying DNA Evidence

Since only a few cells can be sufficient to obtain useful DNA information to help your case, the list
below identifies some common items of evidence that you may need to collect, the possible location
of the DNA on the evidence, and the biological source containing the cells. Remember that just
because you cannot see a stain does not mean there are not enough cells for DNA typing. Further,
DNA does more than just identify the source of the sample; it can place a known individual at a crime
scene, in a home, or in a room where the suspect claimed not to have been. It can refute a claim of
self-defense and put a weapon in the suspect's hand. It can change a story from an alibi to one of
consent. The more officers know how to use DNA, the more powerful a tool it becomes.

Possible Location
Source of
Evidence of DNA on the
DNA
Evidence

 baseball bat or sweat, skin,

handle, end
similar weapon blood, tissue

 hat, bandanna, sweat, hair,

inside
or mask dandruff

nose or ear pieces,

 eyeglasses sweat, skin
lens

mucus, blood,
 facial tissue,
surface area sweat, semen,
cotton swab
ear wax

blood, sweat,
 dirty laundry surface area
semen

 toothpick tips saliva

 used cigarette cigarette butt saliva

 stamp or
licked area saliva
envelope

inside/outside
 tape or ligature skin, sweat
surface

 bottle, can, or
sides, mouthpiece saliva, sweat
glass

inside/outside semen, vaginal

 used condom
surface or rectal cells

sweat, hair,
 blanket, pillow,
surface area semen, urine,
sheet
saliva

 "through and
outside surface blood, tissue
through" bullet

person's skin or
 bite mark saliva
clothing

 fingernail, blood, sweat,

scrapings
partial fingernail tissue

The Nature of DNA

How do we know that genomes are composed of DNA? Using histochemical and physical techniques,
it is relatively simple to demonstrate this fact for eukaryotic nuclear chromosomes. DNA-binding dyes
such as Feulgen or DAPI primarily stain the nuclear chromosomes in cells and to a lesser extent also
stain the mitochondria and chloroplasts. Furthermore if a mass of cells is ground up and its
components fractionated, it becomes clear that the bulk of DNA can be isolated from the nuclear
fraction, and the remainder from mitochondria and chloroplasts.

That DNA is the hereditary material has now been demonstrated in many prokaryotes and
eukaryotes. Cells of one genotype (the recipient) are exposed to DNA extracted from another (the
donor), and donor DNA is taken up by the recipient cells. Occasionally a piece of donor DNA
integrates into the genome of the recipient and changes some aspect of the phenotype of the
recipient into that of the DNA donor. Such a result demonstrates that DNA is indeed the substance
that determines genotype and therefore is the hereditary material

The Three Roles of DNA

Even before the structure of DNA was elucidated, genetic studies clearly indicated several properties
that had to be fulfilled by hereditary material.

One crucial property is that essentially every cell in the body has the same genetic makeup; therefore,
the genetic material must be faithfully duplicated at every cell division. The structural features of DNA
that allow such faithful duplication will be considered later in this chapter.

Secondly, the genetic material must have informational content, since it must encode the constellation
of proteins expressed by an organism. How the coded information in DNA is deciphered into protein
will be the subject of Chapter 3.

Finally, although the structure of DNA must be relatively stable so that organisms can rely on its
encoded information, it must also allow the coded information to change on rare occasion. These
changes, called mutations, provide the raw material—genetic variation—that evolutionary selection
operates on. We will discuss the mechanisms of mutation in Chapter 7.

Properties of DNA

What are the 2 types of DNA?

There are two types of DNA in the cell – autosomal DNA and mitochondrial DNA. Autosomal DNA
(also called nuclear DNA) is packaged into 22 paired chromosomes. In each pair of autosomes, one
was inherited from the mother and one was inherited from the father. Autosomal DNA is passed down
from both the mother and the father and provides clues to a person’s ancestry.

mitochondria are organelles that are responsible for the cell’s energy production. Mitochondria contain
their own DNA called mitochondrial DNA. Mitochondrial DNA has one chromosome that codes for the
specific proteins needed for the metabolic processes that mitochondria perform. Mitochondrial DNA
replicates separately from the rest of the cell and is passed down only from the mother.

Noncoding and coding DNA

Noncoding DNA is sometimes referred to as junk DNA. However, noncoding DNA does have a
purpose. Noncoding DNA typically refers to any DNA that does not code for a protein. This type of
DNA is now referred to as regulatory DNA. The regulatory function of this DNA is to determine when
and where some genes are transcribed. Noncoding DNA also provides chromosomal structure and
binding sites for regulatory proteins. Much research is being conducted on noncoding DNA.

Coding DNA is responsible for harboring the specific DNA sequences that encode instructions for
making proteins.

What are the 3 forms of DNA?

Three major forms of DNA are double stranded and connected by interactions between
complementary base pairs. These are terms A-form, B-form,and Z-form DNA.
B-form DNA
The information from the base composition of DNA, the knowledge of dinucleotide structure, and the
insight that the X-ray crystallography suggested a helical periodicity were combined by Watson and
Crick in 1953 in their proposed model for a double helical structure for DNA. They proposed two
strands of DNA -- each in a right-hand helix -- wound around the same axis. The two strands are held
together by H-bonding between the bases (in anti conformation) as shown in Figure 2.5.1 .

Major groove Major groove

Minor groove Minor groove

Figure 2.5.1 : (left) An A:T base pair and (right) a G:C base pair

Bases fit in the double helical model if pyrimidine on one strand is always paired with purine on the
other. From Chargaff's rules, the two strands will pair A with T and G with C. This pairs a keto base
with an amino base, a purine with a pyrimidine. Two H-bonds can form between A and T, and three
can form between G and C. This third H-bond in the G:C base pair is between the additional exocyclic
amino group on G and the C2 keto group on C. The pyrimidine C2 keto group is not involved in
hydrogen bonding in the A:T base pair.

These are the complementary base pairs. The base-pairing scheme immediately suggests a way to
replicate and copy the the genetic information.

 
Figure 2.5.2 : Antiparallel (a), plectonemically coiled (b, c, d) DNA strands. The arrows in a are
pointed 3’ to 5’, but they illustrate the antiparallel nature of the duplex. The two strands of the duplex
are antiparallel and plectonemically coiled. The nucleotides arrayed in a 5' to 3' orientation on one
strand align with complementary nucleotides in the the 3' to 5' orientation of the opposite strand.

The two strands are not in a simple side-by-side arrangement, which would be called a paranemic
joint (Figure 2.5.3 ). (This will be encountered during recombination in Chapter 8.) Rather the two
strands are coiled around the same helical axis and are intertwined with themselves (which is referred
to as a plectonemic coil). One consequence of this intertwining is that the two strands cannot be
separated without the DNA rotating, one turn of the DNA for every "untwisting" of the two strands.
Figure 2.5.3 : Duplex DNA has the two strands wrapped around each other in a plectonemic coil
(left), not a paranemic duplex (right).

Major and minor groove

The major groove is wider than the minor groove in DNA (Figure 2.5.2𝑑 ), and many sequence
specific proteins interact in the major groove. The N7 and C6 groups of purines and the C4 and C5
groups of pyrimidines face into the major groove, thus they can make specific contacts with amino
acids in DNA-binding proteins. Thus specific amino acids serve as H-bond donors and acceptors to
form H-bonds with specific nucleotides in the DNA. H-bond donors and acceptors are also in the
minor groove, and indeed some proteins bind specifically in the minor groove. Base pairs stack, with
some rotation between them.

A-form nucleic acids and Z-DNA

Three different forms of duplex nucleic acid have been described. The most common form, present in
most DNA at neutral pH and physiological salt concentrations, is B-form. That is the classic, right-
handed double helical structure we have been discussing. A thicker right-handed duplex with a
shorter distance between the base pairs has been described for RNA-DNA duplexes and RNA-RNA
duplexes. This is called A-form nucleic acid.

A third form of duplex DNA has a strikingly different, left-handed helical structure. This Z DNA is
formed by stretches of alternating purines and pyrimidines, e.g. GCGCGC, especially in negatively
supercoiled DNA. A small amount of the DNA in a cell exists in the Z form. It has been tantalizing to
propose that this different structure is involved in some way in regulation of some cellular function,
such as transcription or regulation, but conclusive evidence for or against this proposal is not available
yet.
Differences between A-form and B-form nucleic acid
The major difference between A-form and B-form nucleic acid is in the conformation of the
deoxyribose sugar ring. It is in the C2' endoconformation for B-form, whereas it is in the C3'
endoconformation in A-form. As shown in Figure 2.5.4 , if you consider the plane defined by the C4'-
O-C1' atoms of the deoxyribose, in the C2' endoconformation, the C2' atom is above the plane,
whereas the C3' atom is above the plane in the C3' endoconformation. The latter conformation brings
the 5' and 3' hydroxyls (both esterified to the phosphates linking to the next nucleotides) closer
together than is seen in the C2' endoconfromation (Figure 2.16). Thus the distance between adjacent
nucleotides is reduced by about 1 Angstrom in A-form relative to B-form nucleic acid (Figure 2.5.4 ).

A second major difference between A-form and B-form nucleic acid is the placement of base-pairs
within the duplex. In B-form, the base-pairs are almost centered over the helical axis
(Figure 2.5.42.5.4), but in A-form, they are displaced away from the central axis and closer to the
major groove. The result is a ribbon-like helix with a more open cylindrical core in A-form.
Z-form DNA
Z-DNA is a radically different duplex structure, with the two strands coiling in left-handed helices and a
pronounced zig-zag (hence the name) pattern in the phosphodiester backbone. As previously
mentioned, Z-DNA can form when the DNA is in an alternating purine-pyrimidine sequence such as
GCGCGC, and indeed the G and C nucleotides are in different conformations, leading to the zig-zag
pattern. The big difference is at the G nucleotide. It has the sugar in the C3' endoconformation (like A-
form nucleic acid, and in contrast to B-form DNA) and the guanine base is in the synconformation.
This places the guanine back over the sugar ring, in contrast to the usual anticonformation seen in A-
and B-form nucleic acid. Note that having the base in the anticonformation places it in the position
where it can readily form H-bonds with the complementary base on the opposite strand. The duplex in
Z-DNA has to accomodate the distortion of this G nucleotide in the synconformation. The cytosine in
the adjacent nucleotide of Z-DNA is in the "normal" C2' endo, anticonformation.

Figure 2.5.52.5.5: B-form (left), A-form (middle) and Z-DNA (right). (CC BY-SA 4.0; Mauroesguerroto)

Even classic B-DNA is not completely uniform in its structure. X-ray diffraction analysis of crystals of
duplex oligonucleotides shows that a given sequence will adopt a distinctive structure. These
variations in B-DNA may differ in the propeller twist (between bases within a pair) to optimize base
stacking, or in the 3 ways that 2 successive base pairs can move relative to each other: twist, roll, or
slide.

Guidelines in collection of DNA samples

 Before collection, it must be noted that the biological evidence should be
photographed on its general, medium and close-up or extreme close-up view.
It must be sketched, measured and marked before collecting.
 Requires a wide range of items and biological materials to be collected at the
crime scene. These can be sorted through and analyzed later at the lab to
determine their importance or lack to the case.
 Investigators should make sure not to touch any of the evidence with their
bare hand.
 The person collecting the evidence should wear protective equipment like
shoe cover, hair tie, sterilized rubber gloves and avoid touching their face,
hair, glasses, etc. while collecting.
 Guidelines recommend that while handling and packaging evidence, avoid
sneezing, coughing or even talking, for these reasons, many investigators use
surgical masks when processing the crime scene.
 In packaging items that have apparent wet stains it is necessary to allow
enough time for drying before packaging.
 Always use paper containers (bags, boxes, envelopes) when collecting DNA
samples. Packaging wet items in plastic will moist and creates conditions
favorable for the growth of bacteria and mold and will have deleterious effects
on the sample.
DNA Evidence
Biological Samples for DNA Quantities
Analysis
Whole blood/ Blood stains 1-2 ml or 1 gram
Saliva/ Buccal scrapings 4 cotton buds
Semen/Seminal stains 3 swabs
Urine 100 ml
Hairs with Follicles or Root 2-5 pieces
Muscle tissues/ Cells 15 mg each
Bones and Organs 1-5 grams
Sperm Cells 3 swabs
Dried tissues 100 mg

DNA Profiling (also called DNA testing, DNA typing, or genetic fingerprinting) is a
technique employed by forensic scientists to assist in the identification of individuals
on the basis of their respective DNA profiles. It is especially useful for solving crimes
but can also be used to confirm if the people are related to each other such as for
paternity testing.
USES OF DNA PROFILING
 To prove a crime that has been committed, or established key elements of the
crime
  To link a suspect to the crime or the crime scene
 To establish the identity of persons associated with a crime
 To corroborate victim’s testimony
 To exonerate the innocent
 Negative evidence can help provide proof
 DNA evidence can be more reliable than eyewitness testimony
 The standard method of resolving paternity and immigration disputes
 Identification of dead bodies
 Studying the evolution of human populations
 Studying inherited disorders

Properties of DNA

DNA or deoxyribonucleic acid is the basic hereditary material present in all the cells of an organism
and basically provides a blue print for the cell’s functions, growth, reproduction and death. The
structure of the DNA called the double-stranded helical structure was first described by Watson and
Crick in 1953.

From then enormous progress has been made in synthesis, sequencing, and manipulation of DNA.
The DNA these days can be sequenced or analysed for minute details and even genes may be
inserted to cause changes in the DNA function and structure.

This illustration depicts DNA packed tightly into chromosomes, as well as a DNA molecule unwound
to reveal its 3-D structure. Credit: Darryl Leja, NHGRI

Structure of DNA
The DNA is a polymer molecule with four types of basic chemicals. These are called the
deoxyribonucleotides. They contain:

 a sugar (deoxyribose)
  a negatively charged phosphate group
 the bases:
 adenine (A)
 cytosine (C)
 guanine (G)
 thymine (T)

The nucleotides are linked together by covalent phosphodiester bonds

The Building Blocks of DNA

DNA has three types of chemical component: phosphate, a sugar called deoxyribose, and four
nitrogenous bases—adenine, guanine, cytosine, and thymine. Two of the bases, adenine and
guanine, have a double-ring structure characteristic of a type of chemical called a purine. The other
two bases, cytosine and thymine, have a single-ring structure of a type called a pyrimidine. The
chemical components of DNA are arranged into groups called nucleotides, each composed of a
phosphate group, a deoxyribose sugar molecule, and any one of the four bases. It is convenient to
refer to each nucleotide by the first letter of the name of its base: A, G, C, and T. Figure 2-1 shows the
structures of the four nucleotides in DNA.

Chemical structure of the four nucleotides (two with purine bases and two with pyrim-idine bases) that
are the fundamental building blocks of DNA. The sugar is called deoxyribose because it is a variation
of a common sugar, ribose, which has one more oxygen atom.

DNA Is a Double Helix

DNA is composed of two side-by-side chains (“strands”) of nucleotides twisted into the shape of a
double helix. The two nucleotide strands are held together by weak associations between the bases
of each strand, forming a structure like a spiral staircase (Figure 2-2). The backbone of each strand is
a repeating phosphate–deoxyribose sugar polymer. The sugar-phosphate bonds in this backbone are
called phosphodiester bonds. The attachment of the phosphodiester bonds to the sugar groups is
important in describing the way in which a nucleotide chain is organized. Note that the carbons of the
sugar groups are numbered 1′ through 5′. One part of the phosphodiester bond is between the
phosphate and the 5′ carbon of deoxyribose, and the other is between the phosphate and the 3′
carbon of deoxyribose. Thus, each sugar-phosphate backbone is said to have a 5′-to-3′ polarity, and
understanding this polarity is essential in understanding how DNA fulfills its roles. In the double-
stranded DNA molecule, the two backbones are in opposite, or antiparallel, orientation, as shown in
Figure 2-2. One strand is oriented 5′ → 3′; the other strand, though 5′ → 3′, runs in the opposite
direction, or, looked at another way, is 3′ → 5′.

The arrangement of the components of DNA. A segment of the double helix has been unwound to
show the structures more clearly. (a) An accurate chemical diagram showing the sugar-phosphate
backbone in blue and the hydrogen bonding of bases in the center of the molecule. (b) A simplified
version of the same segment emphasizing the antiparallel arrangement of the nucleotides, which are
represented as L-shaped structures with 5′ phosphate “toes” and 3′ “heels.”
The bases are attached to the 1′ carbon of each deoxyribose sugar in the backbone of each strand.
Interactions between pairs of bases, one from each strand, hold the two strands of the DNA molecule
together. The bases of DNA interact according to a very straightforward rule, namely, that there are
only two types of base pairs: A·T and G·C. The bases in these two base pairs are said to be
complementary. This means that at any “step” of the stairlike double-stranded DNA molecule, the only
base-to-base associations that can exist between the two strands without substantially distorting the
double-stranded DNA molecule are A·T and G·C.

The association of A with T and G with C is through hydrogen bonds. The following is an example of a
hydrogen bond:
Each hydrogen atom in the NH2 group is slightly positive (δ+) because the nitrogen atom tends to
attract the electrons involved in the N–H bond, thereby leaving the hydrogen atom slightly short of
electrons. The oxygen atom has six unbonded electrons in its outer shell, making it slightly negative
(δ−). A hydrogen bond forms between one slightly positive H and one slightly negative atom—in this
example, O. Hydrogen bonds are quite weak (only about 3 percent of the strength of a covalent
bond), but this weakness (as we shall see) is important to the DNA molecule’s role in heredity. One
further important chemical fact: the hydrogen bond is much stronger if the participating atoms are
“pointing at each other” (that is, if their bonds are in alignment), as shown in the sketch.

Although hydrogen bonds are individually weak, the two strands of the DNA molecule are held
together in a relatively stable manner because there are enormous numbers of these bonds. It is
important that the strands be associated through such weak interactions, since they have to be
separated during DNA replication and during transcription into RNA.

The two paired nucleotide strands automatically assume a double-helical configuration (Figure 2-3),
mainly through interaction of the base pairs. The base pairs, which are flat planar structures, stack on
top of one another at the center of the double helix. Stacking (Figure 2-3c) adds to the stability of the
DNA molecule by excluding water molecules from the spaces between the base pairs. The most
stable form that results from base stacking is a double helix with two distinct sizes of grooves running
around in a spiral. These are the major groove and the minor groove, which can be seen in the
models. A single strand of nucleotides has no helical structure; the helical shape of DNA depends
entirely on the pairing and stacking of the bases in antiparallel strands.

Three representations of the DNA double helix.

Structure of DNA FUnction
How does DNA structure fulfill the requirements of a hereditary molecule?
First, duplication. With the antiparallel orientation of the DNA strands, and the rules for proper base
pairing, we can envision how DNA is faithfully duplicated: each strand serves as an
unambiguous template (alignment guide) for the synthesis of its complementary strand. If, for
example, one strand has the base sequence AAGGCTGA (reading in the 5′-to-3′ direction), then we
automatically know that its complementary strand can have only the sequence (in the 3′-to-5′
direction) TTCCGACT. Replication is based on this simple rule. The two DNA strands separate, and
each serves as a template for building a new complementary strand.

An enzyme called DNA polymerase is responsible for building new DNA strands, matching up each
base of the new strand with the proper complement on the old, template strand. Thus, the
complementarity of the DNA strands underlies the entire process of faithful duplication. This process
will be described more fully in Chapter 4.
The second requirement for DNA is that it have informational content. This informational requirement
for DNA is fulfilled by its nucleotide sequence, which acts as a kind of written language. The third
requirement, mutation, is simply the occasional replacement, deletion, or addition of one or more
nucleotide pairs, resulting in a change of the encoded information.

Collection and preservation of DNA

Evidence Collection

Victim service providers, crime scene technicians, nurse examiners, and other medical personnel
should be aware of important issues involved in identifying, collecting, transporting, and storing DNA
evidence. If DNA evidence is not initially identified at the crime scene or on the victim, it may not be
collected, or it may become contaminated or degraded.

To assist in collection, victims of sexual assault should not change clothes, shower, or wash any part
of their body after the assault. Such evidence as semen, saliva, and skin cells may be found on
clothing or bedding, under fingernails, or in the vaginal, anal, or mouth region.

Evidence on or inside a victim’s body should be collected by a physician or sexual assault nurse
examiner. A medical examination should be conducted immediately after the assault to treat any
injuries, test for sexually transmitted diseases, and collect forensic evidence, such as fingernail
scrapings and hair. Typically, the vaginal cavity, mouth, anus, or other parts of the body that may
have come into contact with the assailant are examined.

The examiner should also take a reference sample of blood or saliva from the victim to serve as a
control standard. Reference samples of the victim’s head and pubic hair may also be collected if hair
analysis is required. A control standard is used to compare known DNA from the victim with that of
other DNA evidence found at the crime scene to determine possible suspect(s).

Given the sensitive nature of DNA evidence, victim service providers should always contact crime
laboratory personnel or evidence technicians when procedural collection questions arise.

Questioned or Unknown Samples

Questioned or unknown samples collected from the crime scene can be any biological sample
including: liquid blood or bloodstains, liquid saliva or saliva stains, and liquid semen or dried semen
stains (including from vasectomized males) deposited on virtually any surface; genital/vaginal/cervical
samples collected on swabs or gauze, or as aspirates; rectal/anal swabs; penile swabs; pieces of
tissue/skin; fingernails; plucked and shed hairs (e.g., head, pubic, body); skin cells on drinking
vessels, clothing (e.g., neck collars, waistbands, hat linings); slides containing tissue, semen, etc.;
and liquid urine.

Samples From Unidentified Bodies

Samples collected from unidentified bodies can include: blood, buccal swabs, hairs, bone, teeth,
fingernails, tissues from internal organs (including brain), muscle, and skin.

Reference Samples From Known Individuals

The most common reference samples collected from known individuals are blood, oral/buccal swabs,
and/or plucked hairs (e.g., head, pubic).

Samples to Use When No Conventional Reference Samples Are Available

Other samples that may be considered when individuals are unavailable or are reluctant to provide
samples include clothing where biological fluids may be deposited (e.g., women's panty crotches or
blood-, saliva-, or semen-stained items) and other clothing in close contact with the body where skin
cells may have rubbed off (e.g., collars, waistbands, hats), bedding (with vaginal/semen stains or
rubbed off skin cells), fingernail clippings, cigarette butts, toothbrushes, hairs in razors and
hairbrushes, discarded facial tissues or handkerchiefs with nasal secretions, condoms, gum, feminine
products, pathology paraffin blocks or slides from previous surgery or from autopsy, and teeth.

Reference Samples From Individuals Who Have Been Transfused

If an individual has received transfusions shortly before the collection of a blood sample (e.g.,
homicide victim), the DNA test results may indicate the presence of DNA from two or more sources.
Generally the predominant DNA types reflect the types from the individual. However, other sources of
reference samples for individuals who have received transfusions may need to be collected. These
would include: blood-stained clothing or other material (bedding, etc.) and oral, vaginal, and other
swabs in addition to the items listed above.

Use of Samples From Relatives for Testing

Because a child inherits half of its DNA from each parent, it is possible to use reference samples
collected from close relatives (e.g., biological father, mother, and/or full siblings or the individual's
spouse and their children) to identify or confirm the identity of bodies that have not been identified
through other means. It is also possible to use reference samples collected from close relatives for
comparison to crime scene samples, for example, in missing body cases where a bloodstain or tissue
sample from a possible crime scene can be tested to demonstrate a biological relationship to known
individuals.

Determination of Paternity or Maternity of a Child or Fetus

Aborted fetal tissue can be analyzed for determining paternity, for example, in sexual assault and/or
incest cases where conception occurred. Paternity and/or maternity of a child can be confirmed using
blood or other samples listed above from the child and the alleged parent(s).

Identifying DNA Evidence

Since only a few cells are needed for a useful DNA sample, the list below identifies some areas at the
crime scene or on the victim that may contain valuable DNA evidence. Remember, even though a
stain cannot be seen, there may be enough cells for DNA typing. Furthermore, DNA does more than
just identify the source of the sample; it can place a known individual at a crime scene, in a home, or
in a room where the suspect claimed not to have been. The more victim service providers know about
properly identifying, collecting, and preserving DNA evidence, the more powerful a tool it becomes.
Possible Location of DNA Evidence Source of DNA Bite mark or area licked Saliva Fingernail
scrapings Blood or skin cells Inside or outside surface Semen or skin cells of used condom Blankets,
sheets, pillows, Semen, sweat, hair, or other bed linens or saliva Clothing, including under- Hair,
semen, blood, garments worn during and or sweat after the assault Hat, bandanna, or mask Sweat,
skin cells, hair, or saliva Tissue, washcloth, or Saliva, semen, hair, skin similar item cells, or blood
Cigarette butt; toothpick; or Saliva rim of bottle, can, or glass Dental floss Semen, skin cells, or saliva
Tape or ligature Skin cells, saliva, or hair

Possible Location of DNA Evidence Source of DNA

Bite mark or area licked Saliva

Fingernail scrapings Blood or skin cells

Inside or outside surface of used

condom Semen or skin cells

Blankets, sheets, pillows, or other bed linens Semen, sweat, hair, or saliva

Clothing, including under- garments Hair, semen, blood, or sweat

worn during and after the assault

Hat, bandanna, or mask Sweat, skin cells, hair, or saliva

Tissue, washcloth, or similar item Saliva, semen, hair, skin cells, or blood

Cigarette butt; toothpick; or rim of bottle, Saliva

can, or glass
Semen, skin cells, Semen, skin cells, Semen, skin cells,

Tape or ligature Skin cells, saliva, or hair

Preservation of DNA

Investigators and laboratory personnel should work together to determine the most probative pieces
of evidence and to establish priorities. Although this brochure is not intended as a manual for DNA
evidence collection, every officer should be aware of important issues involved in the identification,
collection, transportation, and storage of DNA evidence. These issues are as important for the first
responding patrol officer as they are for the experienced detective and the crime scene specialist.
Biological material may contain hazardous pathogens such as the human immunodeficiency virus
(HIV) and the hepatitis B virus that can cause potentially lethal diseases. Given the sensitive nature of
DNA evidence, officers should always contact their laboratory personnel or evidence collection
technicians when collection questions arise.

What is DNA Preservation / DNA Banking?

DNA preservation (also known as DNA banking) is the secure preservation and long-term storage of
an individual’s unique genetic material.

Why is it Important?

DNA contains valuable information about each individual, from the color of their hair, eyes and other
physical traits, to their inherited health risks, medical conditions and ancestral roots. Preserving DNA
allows you to capture all of this information forever. Whether you choose to preserve DNA as a
meaningful way to cherish your loved one, or for tracing hereditary health conditions in your family,
DNA banking creates opportunities that would otherwise be lost forever.

Top 5 Reasons to Preserve DNA

Store DNA for Future Testing

Banked DNA lasts indefinitely and can be accessed for testing at any time. A wide range of genetic
tests are currently available for everything from ancestry testing to disease predisposition testing and
more are yet to come.

Document Hereditary Diseases in the Family

75% of all diseases can be traced back to our genetic makeup. Many of the genetic mutations that
cause cancers and other hereditary diseases have been identified. Preserving your family’s DNA is an
important step in tracing the root cause of hereditary diseases and paves the way for targeted
treatments in the future.

Create New Opportunities for Ancestry Testing

Trace your ancestral roots using DNA. As more markers are identified, DNA ancestry testing will
become much more powerful than it is today. DNA banking is a great way to keep a record of your
family’s DNA, opening new doors in this exciting field and creating opportunities that were never
available before.

Prevent DNA Degradation

Without proper purification and preservation techniques, enzymes and contaminants will eventually
break down the DNA in any sample. DNA preservation ensures the long-term viability of a sample for
future testing purposes.

Open Doors for Personalized Medicine

Studying your family’s DNA may assist doctors in determining which treatment plans will work best for
you, and which ones will be ineffective. This technology is already being used to determine the
tolerance, optimal dosage level and effectivity of drugs used in heart disease and cancer patients.

Contamination

Because extremely small samples of DNA can be used as evidence, greater attention to
contamination issues is necessary when identifying, collecting, and preserving DNA evidence. DNA
evidence can be contaminated when DNA from another source gets mixed with DNA relevant to the
case. This can happen when someone sneezes or coughs over the evidence or touches his/her
mouth, nose, or other part of the face and then touches the area that may contain the DNA to be
tested. Because a new DNA technology called "PCR" replicates or copies DNA in the evidence
sample, the introduction of contaminants or other unintended DNA to an evidence sample can be
problematic. With such minute samples of DNA being copied, extra care must be taken to prevent
contamination. If a sample of DNA is submitted for testing, the PCR process will copy whatever DNA
is present in the sample; it cannot distinguish between a suspect's DNA and DNA from another
source.

To avoid contamination of evidence that may contain DNA, always take the following
precautions:

 Wear gloves. Change them often.

 Use disposable instruments or clean them thoroughly before and after handling each sample.
 Avoid touching the area where you believe DNA may exist.
 Avoid talking, sneezing, and coughing over evidence.
 Avoid touching your face, nose, and mouth when collecting and packaging evidence.
 Air-dry evidence thoroughly before packaging.
 Put evidence into new paper bags or envelopes, not into plastic bags. Do not use staples.

Transportation and storage

When transporting and storing evidence that may contain DNA, it is important to keep the evidence
dry and at room temperature. Once the evidence has been secured in paper bags or envelopes, it
should be sealed, labeled, and transported in a way that ensures proper identification of where it was
found and proper chain of custody. Never place evidence that may contain DNA in plastic bags
because plastic bags will retain damaging moisture. Direct sunlight and warmer conditions also may
be harmful to DNA, so avoid keeping evidence in places that may get hot, such as a room or police
car without air conditioning. For long-term storage issues, contact your local laboratory.

Elimination samples

As with fingerprints, the effective use of DNA may require the collection and analysis of elimination
samples. It often is necessary to use elimination samples to determine whether the evidence comes
from the suspect or from someone else. An officer must think ahead to the time of trial and possible
defenses while still at the crime scene. For example, in the case of a residential burglary where the
suspect may have drunk a glass of water at the crime scene, an officer should identify appropriate
people, such as household members, for future elimination sample testing. These samples may be
needed for comparison with the saliva found on the glass to determine whether the saliva is valuable
evidence. In homicide cases, be sure to collect the victim's DNA from the medical examiner at the
autopsy, even if the body is badly decomposed. This may serve to identify an unknown victim or
distinguish between the victim's DNA and other DNA found at the crime scene.

When investigating rape cases, it may be necessary to collect and analyze the DNA of the victim's
recent consensual partners, if any, to eliminate them as potential contributors of DNA suspected to be
from the perpetrator. If this is necessary, it is important to approach the victim with extreme sensitivity
and provide a full explanation of why the request is being made. When possible, the help of a qualified
victim advocate should be enlisted for assistance.

How to Store DNA

DNA is very sensitive and can easily degrade in certain conditions. Thus, proper storage is required to
ensure high experimental standards. There are several mechanisms to store DNA for long periods of
time.

What causes DNA to degrade?

Chemical degradation is the main threat to DNA preservation. Potential nuclease contamination and
the presence of free radicals can also cause damage to DNA and other nucleic acids, such as RNA.
Storage strategies for DNA will depend on the type of DNA, temperature of storage, intended use of
the sample, and the length of time that the DNA is to be stored for.

Storage in vitreous state

In vitreous state, molecules cannot diffuse, this means that a proton’s movement is limited to around
one atomic diameter every 200 years, which effectively inhibits all chemical and nuclease
degradation. Adding moisture to the dry state or raising the temperature will reestablish reactivity and
movement of the protons, once more exposing DNA to potential damage.

DNA stored in a dry state will also remove water, which is active in hydrolytic reactions and can
degrade DNA. Since moisture can reestablish proton movement, DNA stored in the dry state should
be kept at low humidity.

Drying can be done by spray drying, spray freeze drying, and air drying or lyophilization. Of these, the
latter is the cheapest and most popular method. Another method of dry long-term storage is on FTA
cards, which enables preservation for 17 years. In general, DNA storage in dry conditions optimum
ibecause hydrolysis is the biggest cause of DNA degradation.

Medium length storage

Storage of the sample at -20 °C and -80 °C is less effective than storage in vitreous state, but can
provides a useful short-term solution. Liquid nitrogen storage preserves DNA quality over the course
of decades, whereas storage at -20 °C and -80 °C can prevent degradation for months or years.

To prevent degradation by chemical and enzymatic processes, DNA is often stored as a precipitate in
ethanol, at -80 °C. This increases nucleic acids stability, but means that the ethanol must be removed
prior to use. Hence, this technique is considered to be undesirable.

Repeatedly freezing and thawing DNA

A major misconception is that repeated freeze and thaw cycles have a deleterious effect on the quality
of the DNA. However, studies show that repeated freeze and thaw cycles with up to 19 cycles have
no detected DNA degradation.

Some studies indicate that DNA can be satisfactorily kept at room temperature and 4 °C. Such
samples were kept in TE buffer, and were stable for 6 to 12 months. However, such DNA samples
need to be monitored for DNA concentration and evaporation.

Mitochondrial DNA

Mitochondrial DNA is the small circular chromosome found inside mitochondria. The mitochondria are
organelles found in cells that are the sites of energy production. The mitochondria, and thus
mitochondrial DNA, are passed from mother to offspring.
Inside the mitochondrion is a certain type of DNA. That's different in a way from the DNA that's in the
nucleus. This DNA is small and circular. It has only 16,500 or so base pairs in it. And it encodes
different proteins that are specific for the mitochondrial. Now, remember those pathways that are
within the mitochondrion for producing energy. Some of the enzymes in those pathways, and some of
the proteins that are needed to function in those pathways, are produced by the mitochondrial DNA.
The mitochondrial DNA is critically important for many of the pathways that produce energy within the
mitochondria. And if there's a defect in some of those mitochondrial DNA bases, that is to say a
mutation, you will have a mitochondrial disease, which will involve the inability to produce sufficient
energy in things like the muscle and the brain, and the kidney. Mitochondrial DNA, unlike nuclear
DNA, is inherited from the mother, while nuclear DNA is inherited from both parents. So this is very
helpful sometimes in determining how a person has a certain disorder in the family. Sometimes a
disease will be inherited through the mother's line, as opposed to both parents. You can tell from a
pedigree or a group of family history whether or not this is a mitochondrial disease because of that.

William Gahl, M.D., Ph.D.

Mitochondrial Anatomy and Physiology

Mitochondria are spherical, double-membrane-bound organelles that rely on the distinct functions of
their two membranes: the outer mitochondrial membrane and the inner mitochondrial membrane
(Figure 1). The outer mitochondrial membrane contains porins that allow the passage of molecules
smaller than 5 kilodaltons; it also contains a large multiprotein translocase complex that recognizes
mitochondrial signal sequences on larger proteins and permits their passage. The inner mitochondrial
membrane contains all of the components of the electron transport system and the ATP synthase
complex; it also has many invaginations, called cristae, that greatly increase its total surface area. The
double membranes form two mitochondrial compartments: the intermembrane space, located
between the inner and outer mitochondrial membranes, and the matrix, located inside the inner
membrane. Mitochondrial DNA is housed in the mitochondrial matrix.

Mitochondrial vs. Nuclear DNA

Similar to the nuclear genome, the mitochondrial genome is built of double-stranded DNA, and it
encodes genes (Figure 2). However, the mitochondrial genome differs from the nuclear genome in
several ways (Taylor & Turnbull, 2005). Many interesting features distinguish human mitochondrial
DNA from its nuclear counterpart, including the following:

 The mitochondrial genome is circular, whereas the nuclear genome is linear (Figure 3).
 The mitochondrial genome is built of 16,569 DNA base pairs, whereas the nuclear genome is
made of 3.3 billion DNA base pairs.
 The mitochondrial genome contains 37 genes that encode 13 proteins, 22 tRNAs, and 2
rRNAs.
 The 13 mitochondrial gene-encoded proteins all instruct cells to produce protein subunits of
the enzyme complexes of the oxidative phosphorylation system, which enables mitochondria
to act as the powerhouses of our cells.
 The small mitochondrial genome is not able to independently produce all of the proteins
needed for functionality; thus, mitochondria rely heavily on imported nuclear gene products.
 One mitochondrion contains dozens of copies of its mitochondrial genome. In addition,
each cell contains numerous mitochondria. Therefore, a given cell can contain several
thousand copies of its mitochondrial genome, but only one copy of its nuclear genome.

Figure 3

 The mitochondrial genome is not enveloped, and is it not packaged into chromatin.

 The mitochondrial genome contains few, if any, noncoding DNA sequences. (Three percent of
the mitochondrial genome is noncoding DNA, whereas 93% of the nuclear genome is
noncoding DNA).
  Some mitochondrial coding sequences (triplet codons) do not follow
the universal codon usage rules when they are translated into proteins.
 Some mitochondrial nucleotide bases exhibit functional overlap between two genes; in other
words, the same nucleotide can sometimes function as both the last base of one gene and
the first base of the next gene.
 The mitochondrial mode of inheritance is strictly maternal, whereas nuclear genomes are
inherited equally from both parents. Therefore, mitochondria-associated disease mutations
are also always inherited maternally.
 Mitochondrial genes on both DNA strands are transcribed in a polycistronic manner: Large
mitochondrial mRNAs contain the instructions to build many different proteins, which are
encoded one after the next along the mRNA. In contrast, nuclear genes are usually
transcribed one at a time from their own mRNA.

Mitochondrial DNA Mutations

As previously mentioned, one cell contains numerous mitochondria, and each mitochondrion contains
dozens of copies of the mitochondrial genome. Moreover, the mitochondrial genome has a
higher mutation rate (about 100-fold higher) than the nuclear genome. This leads to a
heterogeneous population of mitochondrial

DNA within the same cell, and even within the same mitochondrion; as a result, mitochondria are
considered heteroplasmic. When a cell divides, its mitochondria are partitioned between the two
daughter cells. However, the process of mitochondrial segregation occurs in a random manner and is
much less organized than the highly accurate process
of nuclear chromosome segregation during mitosis. As a result, daughter cells receive similar, but not
identical, copies of their mitochondrial DNA.
Why do mitochondria have such a high mutation rate? A nuclear gene, called DNA
polymerase gamma (POLG), encodes the DNA polymerase responsible for replicating the
mitochondrial genome. The POLG protein consists of two domains: a catalytic domain that exhibits
polymerase activity, and an exonuclease domain that is involved in the recognition and removal of
DNA base-pair mismatches that occur during DNA replication. A recent study suggests that
mitochondria may have a nucleotide imbalance that leads to decreased POLG fidelity and higher
mitochondrial DNA mutation rates (Song et al., 2005).
In the aforementioned study, Song and colleagues (2005) measured the mitochondrial levels of free
deoxynucleotide triphosphates (dNTPs), the building blocks of new DNA strands made during DNA
replication, in tissues from young and old rats. As shown in Table 1, the researchers did not detect
differences in the nucleotide levels in tissues taken from young versus old rats. However, they found
that mitochondrial dNTP levels were highly divergent, and that dGTP was by far the most abundant
nucleotide in the mitochondria of most tissues. In heart muscle and skeletal muscle, for example, they
found that dGTP represented 85% to 91% of the mitochondrial nucleotide pool, whereas dTTP was
present at 0.5%. Similarly, in the brain, dGTP represented 62% of the mitochondrial nucleotide pool,
whereas dTTP constituted only 4%. In the liver, dGTP, dCTP, dATP, and dTTP made up 37%, 51%,
9%, and 3% of the mitochondrial nucleotide pool, respectively. The abundant dGTP levels in
mitochondria were in stark contrast to the dGTP levels in the rest of the cell: In whole rat embryos,
dGTP represented only 10% of the free nucleotide pool.

Clinical Manifestations of Mitochondrial Mutations

Due to the maternal pattern of mitochondrial inheritance, males with a mitochondrial disease are not
considered to be at risk for transmitting the disorder to their offspring. It's important to remember that
there are many mitochondria within a cell, each with its own mtDNA and potential mutations. Thus,
when discussing mitochondrial mutations, it is necessary to think of mutations present across the
entire mitochondrial population rather than in a single mitochondrion. Although mitochondrial
populations are considered heteroplasmic, with variations among the many mtDNA genomes,
mothers can have mitochondrial populations that are homoplasmic for a given mitochondrial mutation;
in this case, the majority of their mitochondrial genome would harbor the mutation. Homoplasmic
mitochondrial mutations will be transmitted to all maternal offspring; however, due to the complex
interplay between the mitochondrial and nuclear genomes, it is often difficult to predict disease
outcomes, even with homoplasmic mitochondrial populations.
What is special about mitochondrial DNA?

Mitochondrial DNA (mtDNA) has many special features such as a high copy number in cell,
maternal inheritance, and a high mutation rate which have made it attractive to scientists from many
fields. ... mtDNA is characterized by the high rate of polymorphisms and mutations.

Our mitochondrial DNA accounts for a small portion of our total DNA. It contains just 37 of the 20,000
to 25,000 protein-coding genes in our body. But it is notably distinct from DNA in the nucleus. Unlike
nuclear DNA, which comes from both parents, mitochondrial DNA comes only from the mother.

Mitochondrial DNA carries characteristics inherited from a mother in both male and female offspring.
Thus, siblings from the same mother have the same mitochondrial DNA. In fact, any two people
will have an identical mitochondrial DNA sequence if they are related by an unbroken maternal
lineage.

Admissibility of DNA Evidence

DNA is a powerful investigative tool because, with the exception of identical twins, no
two people have the same DNA. In other words, the sequence or order of the DNA
building blocks is different in particular regions of the cell, making each person's
DNA unique. Therefore, DNA evidence collected from a crime scene can link a
suspect to a crime or eliminate one from suspicion in the same way that fingerprints
are used. DNA also can identify a victim through the DNA of relatives if a victim's
body cannot be found. For example, if technicians have a biological sample from the
victim, such as a bloodstain left at a crime scene, the DNA taken from that evidence
can be compared with DNA from the victim's biological relatives to determine if the
bloodstain belongs to the victim. When a DNA profile developed from evidence at
one crime scene is compared with a DNA profile developed from evidence found at
another crime scene, they can be linked to each other or to the same perpetrator,
whether the crime was committed locally or in another state.

DNA evidence in the form of saliva, blood, skin tissue, hair, and semen are often
recovered from crime scenes and can be crucial to the investigation of sexual
assaults and other violent crimes. For example, during a sexual assault, biological
evidence such as hair, skin tissue, semen, blood, or saliva can be left on the victim's
body or at the crime scene. In addition, hair and fiber from clothing, carpet, bedding,
or furniture could be transferred to the victim's body during an assault. This evidence
is helpful in proving that there was physical contact between an assailant and a
victim. DNA properly collected from the victim, crime scene, or suspect can be
compared with known samples to place the suspect at the scene of the crime. If
there is no suspect, however, a DNA profile of the crime scene can be entered into
the Federal Bureau of Investigation's (FBI) Combined DNA Index System (CODIS),
which allows agencies to match DNA profiles with other profiles entered into local,
state, and national databases to identify a suspect or link serial crimes.

As with fingerprints, the effective use of DNA as evidence may require the collection
and analysis of elimination samples to determine whether biological evidence came
from a suspect or someone else. When investigating sexual assault or rape cases, it
may be necessary to obtain an elimination sample, such as a blood or saliva sample,
from the victim's relatives or consensual sex partner to account for all of the DNA
found on the victim or at the crime scene.
DNA Evidence: A Brief History

The science of DNA testing was developed in 1985 by British scientist Alec Jeffreys.
Genetic evidence was first tested using his method one year later to solve a double
homicide in England and to link the suspect to other previously unsolved rapes and
murders in the area. In 1987, a Florida rapist became the first criminal defendant in
the United States to be convicted through DNA. Genetic material collected at crime
scenes and preserved in evidence lockers also has become an important factor in
exonerating those who were wrongly convicted of violent crimes.

As DNA became the gold standard for identifying criminal suspects, the FBI and
police departments throughout the U.S. started assembling databases. Additionally,
sex offenders in all states are now required to submit DNA samples to their local
police department. Unfortunately, many crime labs are overwhelmed with backlogs
of genetic samples and may be unable to process them in a timely fashion.

How Does DNA Testing Work?

A sufficient amount of DNA may be found in virtually any type of biological evidence.
For violent crimes, such evidence typically comes from blood or other bodily fluids.
Hair and skin cells left at the crime scene also may provide investigators with enough
DNA for testing purposes.

DNA evidence is analyzed using the polymerase chain reaction (PCR) method,
which allows for very small samples to be tested and identified. Once the sample is
tested, it may be cross-referenced with DNA profiles already in a database or with
genetic data provided by a suspect.

While DNA testing is not completely foolproof, it is more than 99 percent accurate (in
fact, there is only a one in one billion chance that the DNA of two individuals will
match). Typically, errors in testing are the result of mix-ups in the lab or the
contamination of samples. Additionally, each state has specific rules for DNA sample
collection and handling. Courts might not allow the use of genetic evidence in court if
these requirements are not met.

Other Uses of Genetic Evidence

In addition to criminal investigations and trials, DNA can also be used to exonerate
wrongly accused individuals. This is particularly important for those convicted of
serious crimes solely on the basis of eyewitness testimony, which is not always
reliable. More than 250 people have been exonerated through post-conviction DNA
tests, according to the Innocence Project.

Also, DNA can be used to determine paternity in child support cases; to identify the
remains of crime and accident victims; and to conduct genealogical research.

Rules on DNA in the Philippines

In the Philippines rule on DNA evidence was approved in 2007 by former chief
justice Renato Puno.
Section 4. Application for DNA Testing Order. – The appropriate court may, at any time,
either motu proprio or on application of any person who has a legal interest in the matter in
litigation, order a DNA testing. Such order shall issue after due hearing and notice to the
parties upon a showing of the following:

A biological sample exists that is relevant to the case;

The biological sample: (i) was not previously subjected to the type of DNA testing now
requested; or (ii) was previously subjected to DNA testing, but the results may require
confirmation for good reasons;

The DNA testing uses a scientifically valid technique;

The DNA testing has the scientific potential to produce new information that is relevant to
the proper resolution of the case; and

The existence of other factors, if any, which the court may consider as potentially affecting
the accuracy of integrity of the DNA testing.

This Rule shall not preclude a DNA testing, without need of a prior court order, at the behest
of any party, including law enforcement agencies, before a suit or proceeding is commenced.

Section 5. DNA Testing Order.  – If the court finds that the requirements in Section 4 hereof
have been complied with, the court shall –

Order, where appropriate, that biological samples be taken from any person or crime scene
evidence;

Impose reasonable conditions on DNA testing designed to protect the integrity of the
biological sample, the testing process and the reliability of the test results, including the
condition that the DNA test results shall be simultaneously disclosed to parties involved in
the case; and

If the biological sample taken is of such an amount that prevents the conduct of confirmatory
testing by the other or the adverse party and where additional biological samples of the same
kind can no longer be obtained, issue an order requiring all parties to the case or proceedings
to witness the DNA testing to be conducted.

An order granting the DNA testing shall be immediately executory and shall not be
appealable. Any petition for certiorari initiated therefrom shall not, in any way, stay the
implementation thereof, unless a higher court issues an injunctive order. The grant of DNA
testing application shall not be construed as an automatic admission into evidence of any
component of the DNA evidence that may be obtained as a result thereof.

Section 6. Post-conviction DNA Testing. – Post-conviction DNA testing may be available,

without need of prior court order, to the prosecution or any person convicted by final and
executory judgment provided that (a) a biological sample exists, (b) such sample is relevant
to the case, and (c) the testing would probably result in the reversal or modification of the
judgment of conviction.

Section 7. Assessment of probative value of DNA evidence. – In assessing the probative

value of the DNA evidence presented, the court shall consider the following:

The chair of custody, including how the biological samples were collected, how they were
handled, and the possibility of contamination of the samples;

The DNA testing methodology, including the procedure followed in analyzing the samples,
the advantages and disadvantages of the procedure, and compliance with the scientifically
valid standards in conducting the tests;

The forensic DNA laboratory, including accreditation by any reputable standards-setting

institution and the qualification of the analyst who conducted the tests. If the laboratory is not
accredited, the relevant experience of the laboratory in forensic casework and credibility shall
be properly established; and

The reliability of the testing result, as hereinafter provided.

The provisions of the Rules of Court concerning the appreciation of evidence shall apply
suppletorily.

Section 8. Reliability of DNA Testing Methodology. – In evaluating whether the DNA

testing methodology is reliable, the court shall consider the following:

The falsifiability of the principles or methods used, that is, whether the theory or technique
can be and has been tested;

The subjection to peer review and publication of the principles or methods;

The general acceptance of the principles or methods by the relevant scientific community;

The existence and maintenance of standards and controls to ensure the correctness of data
generated;

The existence of an appropriate reference population database; and

The general degree of confidence attributed to mathematical calculations used in comparing

DNA profiles and the significance and limitation of statistical calculations used in comparing
DNA profiles.

Section 9. of DNA Testing Results. – In evaluating the results of DNA testing, the court shall
consider the following:

The evaluation of the weight of matching DNA evidence or the relevance of mismatching
DNA evidence;

The results of the DNA testing in the light of the totality of the other evidence presented in
the case; and that
DNA results that exclude the putative parent from paternity shall be conclusive proof of non-
paternity. If the value of the Probability of Paternity is less than 99.9%, the results of the
DNA testing shall be considered as corroborative evidence. If the value of the Probability of
Paternity is 99.9% or higher there shall be a disputable presumption of paternity.

Section 10. Post-conviction DNA Testing – Remedy if the Results Are Favorable to the
Convict. – The convict or the prosecution may file a petition for a writ of habeas corpus in
the court of origin if the results of the post-conviction DNA testing are favorable to the
convict. In the case the court, after due hearing finds the petition to be meritorious, if shall
reverse or modify the judgment of conviction and order the release of the convict, unless
continued detention is justified for a lawful cause.

A similar petition may be filed either in the Court of Appeals or the Supreme Court, or with
any member of said courts, which may conduct a hearing thereon or remand the petition to
the court of origin and issue the appropriate orders.

Section 11. Confidentiality.  – DNA profiles and all results or other information obtained
from DNA testing shall be confidential. Except upon order of the court, a DNA profile and all
results or other information obtained from DNA testing shall only be released to any of the
following, under such terms and conditions as may be set forth by the court:

Person from whom the sample was taken;

Person from whom the sample was taken;

Lawyers of private complainants in a criminal action;

Duly authorized law enforcement agencies; and

Other persons as determined by the court.

Whoever discloses, utilizes or publishes in any form any information concerning a DNA
profile without the proper court order shall be liable for indirect contempt of the court
wherein such DNA evidence was offered, presented or sought to be offered and presented.

Where the person from whom the biological sample was taken files a written verified request
to the court that allowed the DNA testing for the disclosure of the DNA profile of the person
and all results or other information obtained from the DNA testing, he same may be disclosed
to the persons named in the written verified request.

Section 12. Preservation of DNA Evidence. The trial court shall preserve the DNA evidence
in its totality, including all biological samples, DNA profiles and results or other genetic
information obtained from DNA testing. For this purpose, the court may order the appropriate
government agency to preserve the DNA evidence as follows:

In criminal cases:

for not less than the period of time that any person is under trial for an offense; or
in case the accused is serving sentence, until such time as the accused has served his
sentence;

In all other cases, until such time as the decision in the case where the DNA evidence was
introduced has become final and executory.

The court may allow the physical destruction of a biological sample before the expiration of
the periods set forth above, provided that:

A court order to that effect has been secured; or

The person from whom the DNA sample was obtained has consented in writing to the
disposal of the DNA evidence.

Reference:

https://bio.libretexts.org/Bookshelves/Genetics/Book
%3A_Working_with_Molecular_Genetics_(Hardison)/Unit_I
%3A_Genes_Nucleic_Acids_Genomes_and_Chromosomes/
2%3A_Structures_of_Nucleic_Acids/2.5%3A_B-Form_A-Form_and_Z-
Form_of_DNA

https://nij.ojp.gov/topics/articles/dna-evidence-basics-types-samples-suitable-dna-
testing#:~:text=The%20most%20common%20reference%20samples,e.g.%2C
%20head%2C%20pubic).

https://www.news-medical.net/life-sciences/How-to-Store-DNA.aspx#:~:text=Liquid
%20nitrogen%20storage%20preserves%20DNA,%2C%20at
%20%2D80%20%C2%B0C.

https://www.news-medical.net/life-sciences/How-to-Store-DNA.aspx#:~:text=Liquid
%20nitrogen%20storage%20preserves%20DNA,%2C%20at
%20%2D80%20%C2%B0C.

https://www.news-medical.net/life-sciences/DNA-Properties.aspx

https://www.ncjrs.gov/nij/DNAbro/evi.html

https://www.securigene.com/dna-banking-preservation/understanding-dna-
preservation/

https://www.set.gov.ph/resources/rules-on-dna-evidence/

https://www.findlaw.com/criminal/criminal-procedure/what-is-dna-evidence.html