Learning Objectives

1 Describe the nature of DNA

2 List the various techniques used in DNA profiling

3
Explain how DNA technology is used
in paternity and maternity testing

4
Summarize how DNA technology
has facilitated police work
1
DNA technology

Sex crimes Victim and perpetrator

Rape-murder Only perpetrator left alive

DNA technology Identify the perpetrator

even without witnesses

2
DNA

Human body contains

about 60 trillion cells

60 000 000 000 000

Each cell has a nucleus

Cell
Except red blood cells
3
In the nucleus

Genetic material

Chromosomes,
genes, DNA
The Human Genome

Weighs about 7 picograms

4
DNA

Cannot be found directly at the crime scene

Can find biological materials that contain DNA

Blood, semen, saliva, skin cells,

hair (sometimes) and body parts

5
In the nucleus

Humans

23 pairs of
chromosomes

Chromosomes in
different organisms
6
Humans

23 pairs of chromosomes

22 pairs autosomal Genetic information

2 sex determining chromosomes

XY Male

XX Female
7
Chromosomes

Different sizes & shapes

Chromosomes
8
Chromosomes

Staining with
fluorescent dyes

Distinctive patterns of
bright & dark bands

Different chromosomes
contain different genes Chromosomes
9
Pairs of chromosomes

Homologous

Same size

Contain same genetic structure

One from mother; one from father

10
Genes

DNA material made up of “coding”

and “non-coding” regions

“Coding” regions Genes

“Non-coding” regions 90 to 95% of DNA

Junk DNA
11
Genes

Number of genes in the genome

varies from species to species

Complex organisms More genes

Bacteria Several 100s to 1000s

Humans 30 000

12
What are genes made of?

Made of base pairs

A single gene

1000 to 10 000 base pairs

What is DNA made of?

13
What is DNA made of?

Sugar

Ribose
Ribose

2-deoxyribose

2-deoxyribose
14
What is DNA made of?

- H2O

2-deoxyribose A phosphate ester

of 2-deoxyribose

15
What is DNA made of?

Bases Attached to 2-deoxyribose

Base

Phosphate ester Nucleotide

16
Bases

Heterocyclic

Nitrogen containing

Aromatic

4 bases

17
Bases
NH2 O

N N
N NH
Adenine Guanine
N N N N NH2
H H

NH2 O
H3C
N NH
Cytosine Thymine
N O N O
H H

18
4 possible nucleotides
NH2 O

N N
N NH
O O
N N -O P O N
-O P O N NH2
O O
O- O- H H
A G
H H
H H H H
OH H OH H

NH2 O
H 3C
N NH
O O
-O P O N O -O P O N O
O O

C O-
T
O- H H H H
H H H H
OH H OH H

19
What is DNA? O N
NH2

N
O
P
O N N

A
O
O

4 nucleotides in
O
O
N

polymer chain
O NH
P
O
N N NH2
O

G
O

NH2
O
O

The phosphate
P N
O
O N O

links the sugars C

O
O

O H 3C
NH
O
P
O
O N O
O

T
O

20
What is a double helix?

DNA Two chains wound together

Held in place by hydrogen bonds

Stable under normal conditions

21
Hydrogen bonds

Makes water a liquid

H O H
O H
H

Bases in DNA hydrogen bond together

22
What is a double helix? O
CH3

H H
N
N N
N H

Complementary base pairing

N
T
O
N N

A and T A H
N
H

O N N
C and G N
N
H
O

N N N
H C
H
G
23
What is a double helix?

Human DNA

3 billion base pairs

Paris japonica
Paris japonica
150 billion base pairs
24
The genetic code

Sequence of base pairs

Write the code with A, T, C and G

25
The genetic code

-C-C-T-G-A-G-G-A-G-
Proline Glutamate

3 nucleotides Code for one amino acid

Part of code for a protein

26
The genetic code

-proline-glutamate-glutamate-

Part of the gene for hemoglobin

One wrong letter Different amino acid

Different protein
27
The genetic code

-C-C-T-G-A-G-G-A-G-
-proline-glutamate-glutamate-

-C-C-T-G-T-G-G-A-G-
-proline-valine-glutamate-

Disease Sickle cell anemia

28
Use of DNA

DNA profiling

DNA typing

DNA fingerprinting

Discovered by Professor
Sir Alec Jeffreys Sir Alec Jeffreys
29
Use of DNA

Restriction Fragment Length Polymorphism (RFLP)

Needed large amounts of sample

Use non-coding region of DNA

Why?
30
Use of DNA

Most of what makes us human

beings is common to us all

Few minor differences

31
Use of DNA

Coding region

Not highly individualised

Junk DNA

High variability
32
RFLP

Can use more than one DNA probe

Bill Clinton 7 probes used

More probes Higher probability

33
Gel Electrophoresis & Capillary Gel Electrophoresis

Gel electrophoresis

Analogous to TLC

Capillary gel electrophoresis

Analogous to HPLC

34
DNA Fingerprinting

Required large amounts of samples

Traces are relatively small

Amplify DNA Polymerase Chain

Reaction (PCR)

35
PCR

Each cycle takes

about 5 minutes

Typically run 25 to 35
cycles each time
Thermal cycler

36
PCR results

Allows a specific region of DNA

to be repeatedly duplicated

25 to 35 cycles 1 million copies

Very small trace Converted 1 million times

37
PCR results

Can analyse very small amounts 36 cells

9 cells Low copy number

Saliva from cigarette ends or back of a stamp

Barely visible blood stains

38
Short Tandem Repeats (STR)

Repeating lengths of 3 to 7 base pairs

THO1

AATGAATGAATGAATGAATGAATG…

Varying number of repeats between individuals

39
Short Tandem Repeats (STR)

THO1 6, 8

6 AATG repeated 6 times

8 AATG repeated 8 times

One repeat from each parent

40
Short Tandem Repeats (STR)

Many known STRs

Rely on multiple STRs

Multiplexing

Simultaneous analysis of multiple STRs

41
Short Tandem Repeats (STR)

U.S. 13
STRs Probability
3 1 in 5000
U.K. 10
6 1 in 2 000 000
9 1 in 1 000 000 000
13 1 in 100s of trillions

42
STR & Capillary gel electrophoresis

In pairs One from mother; one from father

43
STR & Capillary gel electrophoresis

Same repeat
number from
both parents

44
Paternity & Maternity Testing

Child

Mother

Father

45
Paternity & Maternity Testing

Child

Mother

Alleged
Father
46
Paternity & Maternity Testing

Same technique to prove sibling relationship

Cannot compare directly between 2 siblings

Inherit different STR repeat

numbers from each parent

47
Sibling relationship

Sibling 1

Sibling 2

Mother

Father
48
Where does your DNA come from?

Half from mother; half from father

Randomly

Siblings will have different DNA

Except identical twins

49
Malaysia, 2009

Identical twins

One arrested for possession of drugs

Police unsure which twin was guilty

Presumption of innocence

50
Other methods

Twin identified by missing tooth

Improving technology

Identical twins do not have 100% identical DNA

Advanced techniques More expensive

51
Marseilles, France

A series of rapes

DNA matched to identical twins

To distinguish the two twins by DNA analysis

Cost 30 million euros

52
DNA technology

Fundamental changes to forensic science

Powerful technique to prove innocence

53
Colin Pitchfork

54
Narborough, England, 1983

Lynda Mann

Raped and murdered

1983

Killer of blood type A

5
5
1986

Second victim Dawn Ashworth

Richard Buckland

Confessed to murder of Dawn Ashworth

Insisted he did not murder Lynda Mann

56
1986

Similarities between the two Same murderer

Alec Jeffreys

Samples from both crime scenes

Sample from Richard Buckland

57
DNA results

Suspect DNA did not match

DNA from the crime scenes

Richard Buckland was innocent

Same man committed both murders

58
1987

DNA screening of local men

Killer was blood type A

Over 1000 men tested

No match
59
Ian Kelly

Gave blood for DNA analysis

under the name of Colin Pitchfork

Reported to the police

60
Colin Pitchfork

Arrested

DNA tested

Matched DNA from

the crime scenes
Colin Pitchfork
61
Colin Pitchfork

Confessed to both murders

Guilty Life in prison

Minimum of 30 years

62
Cold cases

Can be reopened and re-investigated

Evidence properly preserved

DNA databases

63
Cold cases

DNA of known criminals

DNA from old crimes

DNA from newly committed crimes

Check against databases

64
Lesley Molseed

65
Lesley Molseed, 1975

Sexually assaulted

Stabbed to death

Body left out on

the Yorkshire malls

Lesley Molseed
66
Stefan Kiszko

Innocent

Wrongfully imprisoned

16 years
Stefan Kiszko

67
2006

Extracted DNA

Match to Ronald Castree

Charged with the sexual assault

and murder of Lesley Molseed Ronald Castree

68
Problems

1 Most of the original witnesses are dead

Crime occurred over 30 years ago

Ronald Outerridge

69
Problems

Is there proof that the sexual assault and

2
murder were done by the same person?

“To do that it would be necessary to prove that

the semen was deposited during the murder of
the little girl and I found no evidence to show that
was the case. It might have been, it might not."

70
Problems

Lesley Molseed abducted, sexually assaulted,

released, abucted again, and then murdered

Reasonable?

Ronald Castree convicted and sentenced

71
Y o rks hire Ripper

72
1975 onwards

“Yorkshire Ripper”

Murdered prostitutes and some women

1978 Several letters sent to the police

Cassette tape with the Ripper’s voice

73
Cassette tape

Believed to be genuine

Voice on the tape

Strong Geordie accent

Police sent in the wrong direction

74
1981

“Yorkshire Ripper” caught

Peter Sutcliffe

7 attacks and 3 murders later

Tape and letters Hoax

75
2005

Hoaxer case reopened

DNA extract from glue of the envelope

John Humble

Sentenced for perverting the course of justice

76
DNA profiling

Technique first used in the United Kingdom

Spread worldwide

Including the United States

77
Kirk Bloodsworth

1985

Convicted of the rape-

murder of Dawn Hamilton

Kirk Bloodsworth
7
8
1986

Legal technicality Conviction quashed

Re-tried Found guilty again

Verdict confirmed on appeal

Insisted on innocence
79
1993

DNA tests of semen evidence

Did not match Kirk Bloodsworth

Released from prison

80
2003

DNA evidence ran through

the U.S. National Databank

Match to Kimberly Shay Ruffner

81
James Bain

Found guilty of rape in 1974

Based on eyewitness testimony

2009 DNA evidence proved his innocence

Compensated U.S. $1.75 M

82
DNA

What happens if the DNA found at

the crime scene does not match
anybody in the national database?

Other means

83
1988

Lynette White Stabbed to death

Three men convicted

Conviction quashed on appeal

Re-investigation of crime scene

84
2000

Crime scene had been re-decorated

Bloodstains preserved under the paint

DNA sample extracted

No perfect match in the database

85
DNA analysis

Unusual feature in the DNA

600 people shared the feature

70 people shortlisted

1 person stood out

86
DNA analysis

Teenager with a motoring offence

Not him

His uncle

Jeffrey Gafoor Convicted of murder

87
DNA

DNA in the nucleus

Contains our genetic information

88
DNA

Two kinds of DNA within our cell

1 Within the nucleus

2 Within the mitochondria

89
Mitochondrial DNA (mtDNA)

Circular genome 16 569 bases

37 genes

Non-coding regions known as “D-loop”

1 to 3% variation between
unrelated individuals
90
Mitochondrial DNA (mtDNA)

High copy number per cell

More robust than nuclear DNA

More resistant to sample degradation

91
Mitochondrial DNA (mtDNA)

mtDNA inherited strictly from the mother

Not unique to an individual

Faster mutation than nuclear DNA

92
The Romanovs
93
The Romanovs

Russia’s imperial family

1917 Russian revolution

1918

Romanov family murdered / executed

94
The Romanovs

Remains burned and buried

No nuclear DNA

mtDNA extracted

95
Romanov family tree
96
The Romanovs

Intact maternal line

Same mtDNA as Tsar Nicholas

One set of bones identified

97
The Romanovs

Remains of the Tsar’s children

Same mtDNA as the Tsarina

98
Romanov family tree
99
The Romanovs

Prince Philip should have the

same mtDNA as the Tsarina

mtDNA matched the remains

Remains were that of the Romanov family

100
The Romanovs

Rumors

Did the Grand Duchess

Anastasia survive?

Grand Duchess Anastasia

101
Berlin, 1920s

Woman claimed to be the

Grand Duchess Anastasia

Anna Anderson

Died in Virginia, U.S.A. in 1986

Anna Anderson
102
DNA analysis

Tissue from a surgical sample

DNA extracted

Match to Franziska Schanzkowska

103
DNA

All species have DNA

DNA profiling can be done for

any species involved in a crime

104
Maricopa, Arizona, 1992

Woman murdered

Body dumped at an abandoned factory

105
Suspect

Apprehended

Denied having been at the

site where the body was found

Back of his truck Seed pods of the

palo verde tree

106
Suspect

More than one palo verde tree in Arizona

How to prove that the seed pods in

the truck came from the palo verde
tree at the crime scene?

DNA
107
DNA typing

DNA samples of different palo verde trees

Distinguish individual trees

Show and detect variability

DNA from seed pods in the truck matched

the DNA of tree at the crime scene
108
Peter Falconio
& J o a n n e Lees

109
Australia, 2001

Driving around Australia

Northern Territory

Joanne Lees
110
Australia, 2001

Stopped by the driver of another vehicle

Peter Falconio taken out of sight

Shot and never seen again

111
Joanne Lees

Captured

Hands tied with a strap

Escaped and hid in a bush

Strap used to
tie Joanne Lees
Went to the police
112
Police Investigation

Peter Falconio was never found

Found one bloodstain on the road

113
Suspect

Drug smuggler

Bradley Murdoch

Could Bradley Murdoch be

linked to the crime scene?
Bradley Murdoch
114
Police Investigation

vs

Joanne Lees Bradley Murdoch

115
Evidence

1 Blood on her t-shirt

DNA matched to Bradley Murdoch

150 000 000 000 000 000 : 1 of

it being from someone else

Murdoch had had contact with Lees

116
Evidence

2 Gear stick of camper van

DNA matched Bradley Murdoch

Bradley Murdoch was at the crime scene

117
Evidence

3 Strap Low copy number DNA

DNA of Joanne Lees

DNA of Dr. Peter Thatcher

DNA of Bradley Murdoch

100 000 000 : 1

118
Evidence

1 Blood on her t-shirt

Without DNA technology 30 : 1

2 Gear stick of camper van

Without DNA technology Zero

119
Evidence

3 Strap

Without DNA technology

No use as evidence

120
Evidence

“ One of these things by itself might not

have been enough. Put together, there
was a very damning case.” – Rex Wild

Bradley Murdoch convicted

Without DNA technology No case

121
Summary

1 Proving paternity and maternity

2 Police work changed

Very small traces can be individualized

3 DNA databases Solving cold cases

122