Greg Hampikian Presentation

Forensic DNA Analysis
greghampikian@boisestate.edu (208) 781-0438
Katie Monroe
Innocence
project
Jeff Scott Hornoff
Kenneth Wyniemko
Death Penalty Exonerations
Since 1973, 138 people in 25 states have been released
from death row with evidence of their innocence.
Death Penalty Information Center
http://www.deathpenaltyinfo.org/state_by_state
Innocents on Death Row
Average number of years between being sentenced to
death and exoneration: 9.2 years
Number of cases
in which DNA
played a
substantial factor
in establishing
innocence:
*14/122
--Death Penalty Information Center
Charles Fain: on death row for almost
18 years for the rape and murder of a 9year-old girl who was snatched off the
street in Nampa, ID
FOX REPORTERS GANG RAPE
SUSPECTS
BOISE STATE UNIVERSITY LABORATORY
IMPLICATES the FOX 4
• PART ONE
http://www.myfoxatlanta.com/dpp/news
/ITeam_The_GBI_and_DNA_042709
Willie Pete Williams (2007)
“Serial Rapist”
Rapes continued after
arrest
Prosecutor Fred Tokars
denied knowledge
Served 22 years
Sex Assault Kit marked for destruction, supposed to return 1990 to police
Kit found by Cliff Williams, law student
DNA matched other rapes, suspect arrested in Georgia January 2007
On Nov. 29, 1992, Sara Tokars and her sons Ricky, then 7,
and Mikey, then 4, were returning to their suburban
Atlanta home from a Thanksgiving visit with her parents
in Florida when they encountered an armed intruder in
their home.
To some we are the
“Guilty Project”
Twelve days after the initial rape happened in 1985,
there was another rape, which happened within three
miles of the initial rape. Kenneth Wicker was arrested
and convicted in that case. He was sentenced to serve
four years in prison, and served that time.
More than 70% of Exoneration Cases Included Faulty ID
a) Lineups should be presented sequentially (individuals or photos are shown
to the witness one at a time)
b) The individual conducting the photo or live lineup should not know the
identity of the actual suspect. This is called a double-blind procedure
c) Witnesses should be instructed that the suspect may or may not be in the
lineup
d) A minimum of eight photos should be used in photo identification procedures
e) A minimum of six individuals should be used in live identification procedures
f) Witnesses should not receive any feedback during or after the identification
process
g) Witnesses should be asked to give feedback in their own words regarding
their level of confidence in their identification.
How good is DNA at
exonerating?
Crime labs report about 25% of
samples sent by law enforcement
do not match primary suspect
(FBI, GBI, Virginia, Connecticut,
Idaho, Justice Department)
DNA evidence is a frozen moment
in time
Image by collaborator Carl Kriigel, US Army Criminal Investigation Lab
Cells have
Nuclear DNA
and
Mitochondrial
DNA
A Karyotype
Nuclear vs. Mitochondrial DNA
http://www.fbi.gov/hq/lab/fsc/backissu/july1999/dnaf1.htm
Method for obtaining DNA
Sterile Omni-swab for
obtaining…
Images from:
http://www.tritechusa.com/kits/images/FTA%20etc/Sterile-Omni-Swab.jpg
http://faculty.kutztown.edu/friehauf/science_outreach/cells.html
Cheek cells…
Crime Scene Samples &
Reference Samples
• Extract and purify DNA
From bioforensics.com
Differential extraction in
sex assault cases
separates out DNA from
sperm cells
Differential Extraction of Semen Stain
Mild Detergent and enzyme treatment, rupture all epithelial cells, leave
sperm intact
Male Extract
Modified from bioforensics.com
Graphic from Inman & Rudin, An Introduction fo Forensic DNA Analysis. CRC Press.
2 sources of DNA in The Cell
a. Many Mitochondria in Cytoplasm
b. 23 pairs of chromosome in nucleus
cell nucleus
Double stranded
DNA molecule
Target Region for PCR
Individual
nucleotides
www.cstl.nist.gov/biotech/strbase/ppt/4
Vocabulary
• DNA Polymorphism (“many forms”)
Regions of DNA which differ from person to
person
• Locus (plural = loci)
Site or location on a chromosome
• Allele
Different variants which can exist at a locus
• DNA Profile
The combination of alleles for an individual
Modified from www.bioforensics.com
Vocabulary
• Amplification or PCR (Polymerase Chain
Reaction)
A technique for ‘replicating’ DNA in the laboratory
(‘molecular Xeroxing’)
Region to be amplified defined by PRIMERS
Can be ‘color coded’
• Electrophoresis
A technique for separating molecules according to their
size
Modified from www.bioforensics.com
A Old Fashioned
Paternity Print
(RFLP)
DNA
is
Only
part
a
Twins?
Individual
HLA-A
HLA-B
HLA-C
Mother
2,11
44,27
--
Twin 1
2
44,15
3
Twin 2
2,24
44,54
3
PF1
2,3
15
3
13 CODIS Core STR Loci
TPOX
D3S1358
D8S1179
D5S818
FGA
CSF1PO
TH01
VWA
D7S820
AMEL
D13S317
D16S539
www.cstl.nist.gov/biotech/strbase/ppt/4
D18S51
D21S11
AMEL
A Homologous Chromosome Pair
• One from mom
• One from dad
• What is the
difference?
Image: greghampikian@bosiestate.edu
Short Tandem Repeats
(STRs)
AATG
7 repeats
8 repeats
the repeat region is variable between alleles while the
flanking regions where PCR primers bind are constant
Homozygote = both alleles are the same length
Heterozygote = alleles differ and can be resolved from one another
www.cstl.nist.gov/biotech/strbase/ppt/4
PCR Amplification
• DNA regions flanked
by primers are
amplified
•
Groups of amplified STR products are
labeled with different colored dyes
(blue, green, yellow)
DNA Amplification with the
Polymerase Chain Reaction (PCR)
5’
3’
5’
3’
Starting DNA
Template
3’
3’
5’
5’
Separate
strands
(denature)
Forward primer
5’
5’
3’
3’
Make copies
Add
primers
(extend
primers)
5’
(anneal)
3’
3’
5’
Reverse primer
www.cstl.nist.gov/biotech/strbase/ppt/4
PCR Copies DNA Exponentially
through Multiple Thermal Cycles
Original DNA target region
Thermal cycle
In
In32
32cycles
cyclesat
at100%
100%efficiency,
efficiency,1.07
1.07billion
billion
copies
copiesof
oftargeted
targetedDNA
DNAregion
regionare
arecreated
created
ABI Prism 310 Genetic Analyzer
capillary
Syringe with
polymer solution
Injection
electrode
Outlet
buffer
www.cstl.nist.gov/biotech/strbase/ppt/4
Autosampler
tray
Inlet
buffer
Close-up of ABI Prism 310 Sample Loading Area
Electrode
Capillary
Sample Vials
Autosampler Tray
www.cstl.nist.gov/biotech/strbase/ppt/4
ABI 310 Genetic Analyzer:
Capillary Electrophoresis
•Amplified STR DNA
injected onto column
•Electric current applied
•DNA pulled towards the
positive electrode
•DNA separated out by
size:
Large STRs travel slower
•Color of STR detected
and recorded as it
passes the detector
Detector
Window
www.bioforensics.com
Statistical estimates: the product rule
0.222 x 0.222 x 2
= 0.1
www.bioforensics.com
Statistical estimates: the product
rule
1 in 10
x 1 in 111 x 1 in 20
= 0.1
1 in 22,200
x 1 in 14 x 1 in 81
1 in 100
1 in 113,400
1 in 116
x 1 in 17 x 1 in 16
1 in 31,552
1 in 79,531,528,960,000,000
www.bioforensics.com
1 in 80 quadrillion
200th Person Freed By DNA, 2007
http://www.colbertnation.com/the-colbert-report-videos/183230/august-14-2007/jerry-miller
Jerry Miller Served 25 Years for a rape he did not commit
Problems with DNA
Transfer and contamination
Mixture interpretation
Statistical abuse
Jane Durrua, 13, raped and
bludgeoned to death New Jersey, 1968
Joan Conway (left), sister Jane Durrua right
New York Times
Jerry L. Bellamy arrested
DNA Leads to Arrest in '68 Rape and
Murder of Girl, 13
By ROBERT HANLEY
Published: June 17, 2004
DNA evidence that sat in police files for
more than 30 years has led to the arrest of
a 53-year-old sex offender on charges he
raped and murdered a 13-year-old girl who
was walking along a set of railroad tracks
on a November afternoon in 1968.
Off the hook
Authorities said they realized that a lab
scientist had handled DNA evidence
from an Atlantic City case involving
Bellamy and evidence from the Durrua
killing on the same day. That raised the
possibility that Bellamy's DNA had
contaminated the Durrua evidence, and
the prosecutor's office dropped the
charges against him.
New DNA Stains on Death Clothes
NEW JERSEY (WABC) –
March 11, 2008 A convicted killer
already serving a 98-year prison
term for killing a teenage girl was
charged Tuesday with the 1968
murder of a 13-year-old
Middletown girl.
Robert Zarinsky was charged in the
death of Jane Durrua. He died in
prison before trial, 50 years to
the day of his first known murder.
Obituary of Donald Denman
Albuquerque Journal Published on: Fri February 08, 2008
DENMAN -- DONALD (DONNY) EDWARD JR. Donny was born
on November 8, 1954 in Houston, Texas. He was preceded in
death by his parents, Don and Dorothy Denman. He is survived
by his brother, Michael, sisters: Mary, Sherry and Dianna; and
several nieces and nephews. Donny was a free spirit with a
twinkle in his eye and an adventurous personality. He loved to
fish with his brother and mother. Donny was 49 years old when
he left us. We loved him very much and we will miss him each
and every day. A Memorial Service will be held on Saturday,
February 9, 2008 at Immanuel Lutheran Church, 300 Gold
Avenue SE. In lieu of flowers, the family requests that donations
be made to the Salvation Army Alcohol and Drug Rehabilitation
Program in memory of Donny Denman.
FBI Confirms Death with DNA
March 6, 2008
Man's Remains Found in '04 ID'd
It's tough for Michael Denman to look
at a photograph of his brother. As he
stares at the image, he can't help but
wonder.
Would his brother have turned his
life around?
An uncommon correction
May 3rd, 2008
by T. J. Wilham, the Albuquerque Journal.
Donny Denman isn't dead, after all. True, he's
had a funeral. True, a death certificate was
issued in his name. And yes, police have been
looking for his killer ever since they made a DNA
match to some bones found on the West Mesa.
But there was one problem. "I'm very much
alive," he said on a recent Thursday. "It's a trip,
isn't it?"
DNA Transfer
Unlike traditional fingerprints, a defining
feature of DNA fingerprints is their ease
of transfer.
1. Casual Transfer-a fact of life
2. Purposeful Transfer-evidence gathering,
tampering
3. Contamination-laboratory samples with
extraneous profiles
Germany’s Most Lethal Woman
the “Phantom of Heilbronn”
Investigators connected her to six murders, one mysterious
death, and over 40 crime scenes based on DNA swabs from
crime scenes.
This 2007 murder was
believed to be the work of
the phantom killer
April 10, 2008 She robs, she injects herself with heroin, she
seems to kill with almost professional precision – and, as
far as German detectives are concerned, she has no
identity.
For 15 years a mysterious woman has been leaving traces of
her DNA at crime scenes across Europe, suggesting her
involvement in at least six murders and scores of break-ins.
Rarely are there witnesses.
April 10, 2008… The trail began when a boy trod
on a heroin syringe in the sleepy spa town of
Gerolstein, Germany. His parents were so
worried that they insisted on a full blood
analysis. When the DNA traces were fed into a
central data bank a match with genetic material
left at two unsolved murders was found. The first
was of a 63-year-old who was killed in 1993 with
the wire used to bind bouquets. The killer had
left her DNA on the rim of a floral-print tea cup.
April 10, 2008… The woman has left
genetic clues in Germany, France and
Austria,” Chief Inspector Bruno Bösch, the
head of one of three German teams that
has been on the case, said. “I have
travelled 60,000km (37,300 miles) across
Europe, questioning witnesses.”
• April 10, 2008…The evidence suggests that the
Phantom is an addict – hence the desperate
petty burglaries. Flakes of her skin have been
found on a bullet in a gun used between feuding
Romany chiefs. That suggests the Phantom
might be a Gypsy but investigations in the
community have not yielded any results. Nor has
saliva testing of 800 women in the Heilbronn
area.
Police now acknowledge swabs used to collect DNA
samples were contaminated by an innocent woman
working in a factory in Bavaria. March 28, 2009
Birthday Problem and Database
matches
Y Chromosome Testing
• Paternal inheritance.
• Detects male component of a mixture.
• Less discriminating than standard DNA
testing. Statistics = counting method
(linkage).
• Important for detecting the semen donor in
sexual assault mixtures.
Applications of Y-STRs
• Forensic Analysis
– Detect male DNA in a sample containing male
and female DNA (Huge background of female
DNA)
– Aspermic males
– Fingernail Scrapings
– Multiple male donors
– Limits of differential extraction/ tissues
– Gender clarification (amelogenin)
• Paternal Lineage
– Paternity Testing
– Kinship analysis
– Deficiency cases
Nuclear vs. Mitochondrial DNA
http://www.fbi.gov/hq/lab/fsc/backissu/july1999/dnaf1.htm
Dog Mitochondria
DNA Tests Sought in Ga. Child Killings
The Associated Press
Wednesday, November 29, 2006; 12:54 PM
ATLANTA -- Lawyers for Wayne Williams,
blamed for the murders of two dozen children
and young men in the late 1970s and early '80s,
have asked to perform DNA testing on dog hair,
human hair and blood.
Hampikian group mitochondrial projects
Other activities in our group
• Basque mitochondrial heritage study of 95
unrelated families
• Murder of an Alaskan Native Chief