1.

DNA in Forensic

by Dr. Tiana Milanda

 Introduction
 In the late 1800s, efforts to fight crime were
given by photography.
 Alphonse Bertillon was the first to approach
a crime scene with the systematic methods.
He'd capture images at various distances
and take both ground level and overhead
shots.
 Photographs were and are still being used to
record evidence around an crime scene that
then allow forensic scientists and criminal
investigators to have an reference guide for
that particular crime scene.
 Limitations : criminals found may ways to
alter their appearance.

Company Name
 Introduction
 In 1823, John Evangelist Purkinje published his thesis
discussing 9 fingerprint patterns, but he made no mention of
the value of fingerprints for personal identification.
 In 1892, Juan Vucetich made the first criminal
fingerprint identification, because no two fingerprints have
ever been found alike
 They need a huge collections of prints and clerks were
responsible for examining each set of possible prints to find
matches  the process became less tedious and more
realiable with a computer.
 Limitations : fingerprints can be wiped away and gloves can
be worn to keep from leaving fingerprints behind.

Company Name
Introduction
 In 1985, Alec Jeffrey
developed the method of
DNA fingerprinting.

 In the course of his research

on variability in human DNA,
Alec Jeffreys developed a
method of forensic DNA
typing.

 This method, termed DNA

Fingerprinting
 Principles
Two aspects of DNA :
 The genetic variability between individual  The
chemical structure of DNA is the same, but the
order of the base pairs differs

 The unique signature found in each person’s

genetic makeup  DNA fingerprint

 The uniform nature of DNA in a single individual

 every cell in a body shares the same DNA
 DNA Fingerprint
 The basis of genetic testing is that no two
humans DNA is alike (except identical twins).

 Polymorphisms are positions where the

nucleotide sequence is not the same in every
member of the population.
 Human DNA
 In human cells, DNA are packing in nuclear
chromosome and cytoplasmic mitochondria
chromosome
 male
Nuclear
chromosomes
• Human cells : somatic cells
and gametes
• Each somatic cells have
two sets of 23 linear
chromosomes (diploid), 23
maternal chromosomes
and 23 paternal female
chromosomes
homologous pairs
 LOCUS AND ALLELE
Allele:
Refers to the type of DNA.

Locus or Loci:
Refers to the location on
the chromosome

2 alleles per locus

 male

Somatic Cells
• Chromosomes 1-22 
autosomes, chromosomes
23  sex chromosomes,
consisting of X and Y
chromosome
• Autosomes contain genes female
that affects other body
features unrelated to sex
• Sex chromosome (X and
Y) contain genes that
influence sex traits and
the development of
reproductive organs
 Autosomes : coding and non
coding sequences
 Nuclear (and mitochondrial) chromosomes consist of two
types of nucleotides
1. Those that make up the genes, called coding
sequences
2. Those whose function is largerly unknown, called non
coding regions
 The nucleotide in coding and non coding portion are
alike, they differ solely in whether or not they contribute
to one or more of the individual’s trait (phenoptype)
 Nuclear : Non coding sequence

Intergenic DNA
3’ 5’
5’ Gene 1 Gene 2
3’

3’ 5’
5’ Gene 1 Gene 2
3’

Introns
Nucleus :
Eukaryotic genomes full of repeated DNA

Repeat Unit # Repeats

Satellite DNA 100 – 1000 bp ???

Minisatellite DNA or
7 – 100 bp 7 - 80
Variable Number of
Tandem Repeats (VNTR)

Microsatellite DNA or
2 – 6 bp 5 - >40
Short Tandem Repeats
 VNTR
 Rectangle blocks represent repeated DNA sequences at
a particular VNTR location
 Repeats are tandem – clustered together and oriented in
same direction
 Repeats can be removed or added leading to alleles with
different numbers of repeats.
 Types of STRs

Dinucleotide, trinucleotide, tetranucleotide,

pentanucleotide, hexanucleotide.

Simple repeats: identical length and sequence.

Compound repeats: two or more adjacent simple
repeats.
Complex repeats: repeat blocks varying in unit
length and sequence
 Short Tandem Repeats

Person 1 ..GCCAGCTAGCTAGCTAGCTAGCTAGCTTTCAT..
1 2 3 4 5 6

Person 2 ..GCCAGCTAGCTAGCTAGCTAGCTTTCAT..
1 2 3 4 5
Person 3 ..GCCAGCTAGCTAGCTAGCTAGCTAGCTAGCTT..
1 2 3 4 5 6 7

On average, occur every 10,000 nucleotides.

 LOCUS AND ALLELE
Allele:
Refers to the type of DNA.

Locus or Loci:
Refers to the location on
the chromosome

2 alleles per locus

Person 1..GCCAGCTAGCTAGCTAGCTTTCAT..
1 2 3 4
..GCCAGCTAGCTAGCTAGCTAGCTAGCTTTCAT..
1 2 3 4 5 6
Person 2..GCCAGCTAGCTAGCTAGCTAGCTAGCTTTCAT..
1 2 3 4 5 6
..GCCAGCTAGCTAGCTAGCTAGCTAGCTAGCTTTCAT..
1 2 3 4 5 6 7
Person 3..GCCAGCTAGCTAGCTAGCTAGCTTTCAT..
1 2 3 4 5
..GCCAGCTAGCTAGCTAGCTAGCTTTCAT..
1 2 3 4 5

Homozygote = both alleles are the same length

Heterozygote = alleles differ and can be resolved from one
another
 Loci Numenclature

Intergenic DNA
3’ 5’
5’ Gene 1 Gene 2
3’

3’ 5’
5’ Gene 1 Gene 2
3’

Introns
 Loci Nomenclature
• Introns: STR name based on gene.
TH01 - located in the 1st intron (01) of gene for
tyrosine hydroxylase

• Intergenic DNA: based on chromosome

D = DNA
D5S818 5 = chromosome 5
S = single copy in genome
818 = 818th locus
 LOCUS AND ALLELE
Allele:
Refers to the type of DNA.

Locus or Loci:
Refers to the location on
the chromosome

2 alleles per locus

 male

Somatic Cells
• Chromosomes 1-22 
autosomes, chromosomes
23  sex chromosomes,
consisting of X and Y
chromosome
• Autosomes contain genes female
that affects other body
features unrelated to sex
• Sex chromosome (X and
Y) contain genes that
influence sex traits and
the development of
reproductive organs
 male

Somatic Cells
• Human males have an X
and a Y chromosomes
(XY) and females have a
pair of X chromosomes
(XX)
• A set genes found on the female
Y chromosomes, called
SRY (sex-determining
region) determine an
embryo into a male
 Y Chromosome Structure
SRY ~60 Mb total DNA sequence (only
chromosome 22 is smaller)
p AMEL

~2.5 Mb on tips recombine with X

(pseudoautosomal regions)

q 35-36 Mb euchromatin 9.5 Mb

sequenced (27%)

heterochromatin
Genetic variation at
multiple points along the Y
chromosome is combined
to form a Y haplotype for a
Nucleic Acids Res. 28(2), e8 (2000) sample
 Human DNA
 In human cells, DNA are packing in nuclear
chromosome and cytoplasmic mitochondria
chromosome
 Mitochondrial DNA
Mitochondria - The powerhouse of the cell.
The number of mitochondria varies greatly with the type of
cell and stage of its development (ranging 200-1,000)

Mitochondria have
Mitochondria their own DNA
 Mitochondrial DNA

Double Helix Nuclear Mitochondrial DNA

Chromosomes
Each mitochondrion contains two or three circular DNA
Mitochondrial DNA is
only 16,569 letters long
[compared to 3 billion in
nuclear DNA]
 Coding and non coding sequences
 In a mitochondrial chromosome, the coding and non
coding areas are entire separate. Non coding portion
being located in a region reffered as the control region
(also called displacement loop or D-loop)
 The control region contains 1,100 bp and is devided into 2
distinct sections, hypervarible 1 (HV1) and hypervarible 2
(HV2)
History of DNA Testing (Soon)
 DNA fingerprint :
- RFLP Multilocus (fingerprint)
- RFLP single locus
 DNA profiling :
- PCR STR
 Y chromosome testing
 SNP testing
 Mitochondrial DNA testing
 Some uses of DNA
Fingerprint/Profiling
 Forensic work on crime scenes
 Parentage testing
 Victim identification in mass disasters
 Non human DNA analysis
Forensic work on
crime scenes
The Narborough Village Murder
 DNA evidence was compared to DNA samples from
5,500 suspect’s blood by RFLP multilocus  the DNA
did not match.
 A man had given a sample in the name of friend  an
exact match
 Limitation : unless there is a known sample to be used
as a comparison, identity cannot be established
Blood from
Defendant’s defendant’s clothes Victim’s
blood blood
 The Forest Hill Rapist
 In 1988, Victor Lopez was tried for the sexual
assault of three women. Oddly, all three women
had described their assailant as a black man.
Lopez was not black.
 The accused’s blood was analyzed and
compared with sperm left at the sceene  the
DNA was a match  Lopez was found guilty of
the attacks.
 DNA evidence is especially valuable when used
to expose faulty eye-witness testimony
 Simpson/Goldman murders
 In 1994, Nicole Brown Simpson and
Ronald Goldman were murdered
and Nicole’s ex-husband, O.J.
Simpson was a suspect.
 The DNA collected at the crime
scene matched that of O.J.
Simpson.
 Defense lawyers attacked the
procedures used in
collecting,labelling and testing the
evidence
 As a result, the DNA evidence was
not effective, O.J. Simpson was
found not guilty
 Humans error and
sources of
contamination
The great threats to DNA evidence is :
 Human error  the chain of custody requires that the
collection of evidence must be systematically recorded and
access to the evidence must be controlled. The process of the
samples adhere to high standard.
 Damage during the analysis itself  defined standard of
laboratory practice and procedure can help guard againts
errors during DNA forensics analysis
 Band shifting occurs during electrophoresis, when the DNA
fragments in one lane migrates more rapidly than identical
fragments in second line  minor inconsistencies in the gel or
a surplus DNA in one of the lanes
Paternity Testing
Victim identification
in mass disasters
 Victim Identification : September 11
• Nearly 3000 people died as a result of the terrorist attacks
on September 11th, 2001
• Dangerous conditions at ground zero made it very difficult
for forensic scientists to determine the identity of victims
based on their remains.
• Most remains were small bone fragments and teeth
• Collection points were set up around New York City where
family members could make missing person reports as well
as donate items that may contain DNA of the victims.
M-FISys
Mass Fatality Identification System
• Software system created in the wake of September
11th attacks
• First software to combine STR, mtDNA, and SNP in
an integrated matter.
• Out of the 3000 people who died, only about 1700
were identified.
• Can be modified to be used in other disasters or in
natural disasters
• Altered, and used to identify over 800 individuals in
three months following the South Asian Tsunami
Non Human DNA
Analysis
 Non human DNA analysis
 Many legal questions have been answered by the
genetic profiles of plants and animals.
 For example, the two type of ginseng (American ginseng
and Asian ginseng) look almost identical, but they have
very different reputations. Asian ginseng purportedly
boosts energy, whereas American ginseng is prized for
its ability to calm nerves  using DNA sequencing to
help make distinction between two varieties.
 Another example :
- Distinct the varieties derived crossbreeding are
inherently less valuable as wine grapes.
- DNA profiling is regularly used by wildlife
management authorities.
 “Paternity” of Plants

Ancestry of Cabinet Sauvignon grapes

 Wine purists believe hybrid grapes are inferior
 Scientists can examine the DNA cabinet sauvignon
plants
 Found two other varieties : Cabernet Franc & Sauvignon
Blanc
 This challenged the idea that the varieties of these plants
that come from cross-breeding are inherently less
valuable as wine grapes.
Animal Profiling
 Evidence has been used to form
genetic profiles of animals
 Example Is a case in
Pennsylvania
 DNA fingerprinting was used to
prove that a hunter had illegally
killed a bear.
 Authorities collected blood
samples from the bear den and
then compared it with samples
from the check station
 Because of this the DNA analysis
investigators were able to find the
hunter guilty.