Issues in Biotechnology:
The Way We Work With Life
Dr. Albert P. Kausch
life edu.org
Part II of the Course
The Applications of
Biotechnology
A Sweeping General Survey on Life and Biotechnology
The University of Rhode Island
© life_edu
Issues in Biotechnology:
The Way We Work With Life
Dr. Albert P. Kausch
life edu.org
Forensics
25. Trace Evidence
26. DNA based Forensic
27. The National Debate Public Safety vs The Right to Privacy
A Sweeping General Survey on Life and Biotechnology
The University of Rhode Island
Forensics
Lectures 25, 26 & 27
Issues in Biotechnology:
The Way We Work With Life
Dr. Albert P. Kausch
life edu.org
Forensics
DNA Based Evidence
The University of Rhode Island
Forensics:
Bioweapons
DNA Based Forensic Evidence
Forensic DNA Databases: The National Debate
Biotechnology Stocks Project
Time to cash in (or out) as the case may be!!!
$100,000.00!!!!!!!
What happened to your
Invested in Biotech. Stocks this Semester?
1. Select and Research five Biotech companies
2. Print out the current stock quote and annual chart
3. Invest chosen amounts in each. Calculate shares in each.
4. Monitor Stock
5. Print out the stock quote and annual chart Weds.
6. Calculate gains and losses. Submit report.
Are you here?
A. Yes
B. No
C. Uncertain
60
40
20
0
1
2
3
Whan that Aprill, with her shoures soote
The droghte of March is perced to the roote
And bathed every veyne in swich licour,
Of which vertu engendred is the flour;
Whan Zephirus eek with his sweete breeth
Inspired hath in every holt and heeth
The tendre croppes, and the yonge sonne
Hath in the Ram his halfe cours yronne,
And smale foweles maken melodye,
And slepen al the nyght with open eye(So priketh hem Nature in hir corages);
Thanne longen folk to goon on pilgrimages
And especially from every shires ende
Of URI, to 271 Chaffee they wende,
When in April the sweet showers fall
That pierce March's drought to the root
And bathed every vein in sweet liquor
That has power to generate therein and sire the flower;
When Zephyr also has with his sweet breath
,Filled again, in every holt and heath,
The tender shoots and leaves, and the young sun
His half-course in the sign of the Ram has run,
And many little birds make melody
That sleep through all the night with open eye
(So Nature pricks them on to ramp and rage)
Then folk do long to go on pilgrimages
And especially from every shire's end
Of URI they to 271 Chaffee they went
Interpreting the Human Genome
What do we have?
EAKANDWEARYOVERMANYAQUAINTANDCURIOUSVOLUMEOFFOR
GOTTENLOREWHILEINODDEDNEARLYNAPPINGSUDDENLYTHEREC
AMEATAPPINGASOFSOMEONEGENTLYRAPPINGRAPPINGATMYCH
AMBERDOORTISSOMEVISITORIMUTTEREDTAPPINGATMYCHAMO
NCEUPONAMIDNIGHTDREARYWHILEIPONDEREDWBERDOORONLY
THISANDNOTHINGMOREAHDISTINCTLYIREMEMBERITWASINTHEB
LEAKDECEMBERANDEACHSEPARATEDYINGEMBERWROUGHTITSG
HOSTUPONTHEFLOOREAGERLYIWISHEDTHEMORROWVAINLYIHA
DSOUGHTTOBORROWFROMMYBOOKSSURCEASEOFSORROWSORR
OWFORTHELOSTLENOREFORTHERAREANDRADIANTMAIDENWHO
MTHEANGELSNAMELENORENAMELESSHEREFOREVERMOREANDT
HESILKENSADUNCERTAINRUSTLINGOFEACHPURPLECURTAINTHR
ILLEDMEFILLEDMEWITHFANTASTICTERRORSNEVERFELTBEFORE
SOTHATNOWTOSTILLTHEBEATINGOFMYHEARTISTOODREPEATIN
GTISSOMEVISITORENTREATINGENTRANCEATMYCHAMBERDOORS
OMELATEVISITORENTREATINGENTRANCEATMYCHAMBERDOORT
HISITISANDNOTHINGMORE
Interpreting the Human Genome
Organization and structure is needed!
Once upon a midnight dreary, while I pondered, weak and weary,
Over many a quaint and curious volume of forgotten lore,
While I nodded, nearly napping, suddenly there came a tapping,
As of some one gently rapping, rapping at my chamber door.
“Tis some visitor," I muttered, "tapping at my chamber door—
Only this, and nothing more."
Ah, distinctly I remember it was in the bleak December,
And each separate dying ember wrought its ghost upon the floor.
Eagerly I wished the morrow;--vainly I had sought to borrow
From my books surcease of sorrow--sorrow for the lostLenore—
For the rare and radiant maiden whom the angels name
Lenore--Nameless here for evermore.
Pharmacogenomics
Making Sense
Organization and structure is needed!
Once upon a midnight dreary, while I pondered, weak and weary,
Over many a quaint and curious volume of forgotten lore,
While I nodded, nearly napping, suddenly there came a tapping,
As of some one gently rapping, rapping at my chamber door.
“Tis some visitor," I muttered, "tapping at my chamber door—
Only this, and nothing more."
Ah, distinctly I remember it was in the bleak December,
And each separate dying ember wrought its ghost upon the floor.
Eagerly I wished the morrow;--vainly I had sought to borrow
From my books surcease of sorrow--sorrow for the lostLenore—
For the rare and radiant maiden whom the angels name
Lenore--Nameless here for evermore.
Do you know how to make a
nuclear bomb?
A.
B.
C.
D.
Yes
No
Uncertain
I could find out
30
20
10
0
1
2
3
4
Synthetic Biology
A Larger Threat to World Peace than Nuclear War?
To whom much is given
much is required Luke 12:48
Once you know, you cannot unknow Kausch, The Abandon
In much wisdom there is grief
in much knowledge there is pain Ecclesiastes 1:18
Now that you know, know that you now Kausch, The Abandon
Issues in Biotechnology:
The Way We Work With Life
Dr. Albert P. Kausch
life edu.org
Forensics
DNA Based Evidence
The University of Rhode Island
Forensics:
DNA Based Evidence
Forensic DNA Databases: The National Debate
Trace Evidence
I typically watch TV:
A.
B.
C.
D.
E.
0-2 hrs/day
2-3 hrs/day
3-5 hrs/day
5-10 hrs/day
Over 10 hrs/day
50
40
30
20
10
0
1
2
3
4
5
The average American
watches TV:
A.
B.
C.
D.
E.
0-2 hrs/day
2-3 hrs/day
3-5 hrs/day
5-10 hrs/day
Over 10 hrs/day
I typically watch TV:
4 hrs X 365 = 1460 hrs/yr
1460 hrs/yr ÷ 16 hrs waking hrs/day = 91.25 days/yr
91.25 days/yr over 50 yrs = 4562.5 days or 12.5 yrs
Does watching TV
influence teenage sexual
behavior?
Does watching TV
influence teenage violent
behavior?
Children And TV Violence 2012
Hundreds of studies on the effects of TV violence on
children and teenagers have found that children may:
•become "immune" or numb to the horror of violence
•gradually accept violence as a way to solve problems
•imitate the violence they observe on television; and
•identify with certain characters, victims and/or victimizers
Children And TV Violence 2008
•Nearly 2 out of 3 TV programs contain some violence,
averaging about 6 violent acts per hour.
•The average child who watches 2 hours of cartoons a day
may see nearly 10,000 violent incidents each year, of which
the researchers estimate that at least 500 pose a high risk for
learning and imitating aggression and becoming
desensitized to violence.
Center for Communication and Social Policy, University
of California, Santa Barbara (UCSB), National Television
Violence Study, Executive Summary, Volume 3 2008.
Longitudinal Relations Between Children’s Exposure to TV Violence
and Their Aggressive and Violent Behavior in Young Adulthood:
1977–1992
L. Rowell Huesmann, Jessica Moise-Titus, Cheryl-Lynn Podolski, and Leonard D. Eron
University of Michigan
Developmental Psychology 2003 The American Psychological Association, Inc.
2003, Vol. 39, No. 2, 201–221
ABSTRACT
Although the relation between TV-violence viewing and aggression in childhood has been clearly
demonstrated, only a few studies have examined this relation from childhood to adulthood, and these
studies of children growing up in the 1960s reported significant relations only for boys. The current
study examines the longitudinal relations between TV-violence viewing at ages 6 to 10 and adult
aggressive behavior about 15 years later for a sample growing up in the 1970s and 1980s. Follow-up
archival data (N 450) and interview data (N 329) reveal that childhood exposure to media violence
predicts young adult aggressive behavior for both males and females. Identification with aggressive TV
characters and perceived realism of TV violence also predict later aggression. These relations persist
even when the effects of socioeconomic status, intellectual ability, and a variety of parenting factors are
controlled.
Connecticut State
Forensic Science
Laboratory
Michael Adamowicz, Criminalist
Carll Ladd, Lead Criminalist
Forensic Biology
The Rhode Island
State Crime Lab:
Forensic Examinations
Dennis Hilliard, Director
Amy Duhaime
Criminalist III
Rhode Island State Crime Lab
Forensic DNA Testing is
Nothing
Just
ing like CSI!
Forensic Science:
the application of natural
sciences to matters of
the law.
Physical Evidence Analysis
• Is concerned with the recognition,
identification, comparison,
individualization, interpretation and
reconstruction of evidence.
Criminalistics:
Study and evaluate the recognition,
identification, individualization, and
evaluation of physical evidence using
the methods of the natural sciences in
matters of legal significance.
Physical evidence examination can :






Link a suspect with the victim
Link a person to a crime scene
Link an object to a crime
Disprove or support witness testimony
Identify a person
Aid in the Reconstruction of a crime
Chain of Custody
The chain of custody begins when the
evidence is located at the scene even before it
is collected and does not end, until the case
has been adjudicated in court and all appeals
have been exhausted.
Forensic Science Lab Services
Criminalistics
• Forensic Biology
Serology
DNA
• Trace Analysis
• Chemistry
• Instrumentation
Identification
 Latent Print Section
 Questioned
Documents
 Imprints
 Firearms
 Toolmarks
 Forensic Photography
*Crime Scene Reconstruction
How Does DNA Forensic Testing
Help
Help an Investigation?
By Providing Important
Linkages:
 Link suspect to victim
 Link suspect to scene
 Link victim to scene
Forensic DNA Evidence
Circumstantial Evidence
Was a person there?
Patterns
Patterns?
Patterns are the basis of DNA Identification
DNA Profiles, Marker D10S28
C V D E1 E2 E3
C
Patterns in DNA markers can link
a suspect to a crime scene
C = Control
V = Victim
D = Defendant
E = Evidentiary sample
Issues in Biotechnology
If you flip a coin six times and get heads on all six flips, what
is the probability of getting heads on the next toss?
A
B
C
D
F
1/2
1/100
1/1000
1/10,000
1/1,000,000
40
30
20
10
0
1
2
3
4
5
Murder at Rodman Dam, 1988
• In July 1987 Randall Scott Jones and Chris
Reesh, both in their teens, went target shooting
with a 30/30 hunting rifle at the Rodman Dam
Recreation Area in Florida. While they were
shooting, Jone’s pickup truck became stuck in a
sand pit. A fisherman suggested they ask a
couple in a pickup parked nearby for help.
Jones and Reesh approached the truck, where
Kelly Lynn Perry and her fiancé Matthew Brock
were sleeping. The two men debated whether
or not to wake them to ask for assistance.
Murder at Rodman Dam, 1988
• The following morning, fishermen found the
bodies of Perry and Brock in the woods adjacent
to the recreation area. Police investigation
revealed that they had been shot with a 30
caliber bullet and Perry had been sexually
assaulted. Their pickup was reported stolen.
Murder at Rodman Dam, 1988
• In August, Jones was arrested in Mississippi,
found driving Brocks pickup. Reesh was
arrested the next day in Palatka, Florida, after
Jones told police that they were together that
night in July. Both were indicted on counts of
first degree assault and sexual battery.
Murder at Rodman Dam, 1988
• A semen sample E(vs) retrieved from
Perry’s body, and blood samples from
Reesh, S1, and Jones, S2, were
compared at a laboratory that specialized
in DNA testing. The resulting DNA
evidence indicated which man was guilty
of rape.
Murder at Rodman Dam, 1988
• DNA results
Who is most
likely guilty of
the rape?
A. Chris
Reesh
B. Randall
Jones
Murder at Rodman Dam, 1988
• Using the DNA results and other evidence, officials
identified Jones as the rapist and were able to piece
together the events of the crime.
•
Without waking the couple in the pickup, Jones
shot both Perry and Brock in the head at close
range. He and Reesh then dragged the bodies into
the woods nearby. They towed Jone’s truck with
Brock’s pickup and left with both trucks.
•
Later, Jones returned to the crime scene, moved
the bodies further into the woods, and raped Perry.
Murder at Rodman Dam, 1988
• A representative from the DNA lab testified that
the chance of another person having the same
DNA fingerprint as Jones was one in
9,390,000,000, about twice the earth’s
population.
• After deliberating only 15 minutes, the jury
convicted Jones of murder and rape. The judge
sentenced him to a double death sentence,
making this the first case involving DNA
evidence in the U.S. legal history in which the
death sentence was handed down. Reesh was
sentenced to six years in prison and twenty
years probation.
DNA
Uses for DNA Analysis
 Criminal Investigations
 Paternity Cases
 Genetic Disease Diagnosis
 Identifying Endangered Animals
 Identifying Remains from War
 Identifying Accident Victims
DNA
Hereditary material of all living organisms.
Found predominantly in the cell nucleus.
Organized into chromosomes.
Humans-46 chromosomes.
23 maternal & 23 paternal.
Polymer-individual units called nucleotides.
Structure-double helix.
Watson and Crick, 1953.
Forensic Identification:
Basic Principles
Each of us is genetically unique.
If enough genetic variation is tested, each
of us can be uniquely identified.
DNA is found in nearly all cells (blood,
semen, hair, etc.).
DNA from an evidentiary sample can be
matched with DNA from a suspect to
implicate or exonerate.
DNA Casework
1. Forensic Analysis (Criminal).
-132 labs conducting DNA analysis in 49 states.
~ 40,000 cases/year received.
~25,000 analyzed.
~80% sexual assaults.
2. ~30% of the time the suspect is excluded by
DNA.
3. ~ 300,000 paternity cases per year.
Sources of Biological Evidence
•
•
•
•
•
•
•
•
Blood
Semen
Saliva
Urine
Hair
Teeth
Bone
Tissue
Other Possible items for DNA Testing:
1. cigarette butts
2. gloves, bandanas, ski masks, baseball caps
general clothing
3. condoms (inside vs. outside)
4. stains on furniture, pillows, sheets
5. hair clips, lipsticks
6. letters, envelopes, and stamps
7. plant and animal sources of evidence
PCR
Gel Electrophoresis:
the separation of molecules,
DNA, RNA and proteins
by charge and size
Electro refers to the energy of
electricity. Phoresis, from the
Greek verb phoros, means
"to carry across." Thus, gel
electrophoresis refers to the
technique in which molecules
are forced across a span of gel,
motivated by an electrical
current.
The gel matrix acts as a sieve for DNA molecules. Large
molecules have difficulty getting through the holes in the
matrix. Small molecules move easily through the holes.
Because of this, large fragments will lag behind small
fragments as DNA migrates through the gel.
As the separation process continues, the
separation between the larger and smaller
fragments increases.
•Molecular weight markers are often
electrophoresed with DNA.
•Molecular weight markers are usually a mixture of
DNAs with known molecular weights
•Molecular weight markers are used to estimate the
sizes of DNA fragments in a DNA sample
What are some of the DNA technologies
used in forensic investigations?
Restriction Fragment Length Polymorphism (RFLP)
PCR Analysis
STR Analysis
Mitochondrial DNA Analysis
Y-Chromosome Analysis
Restriction Fragment Length Polymorphism
(RFLP)
RFLP is a technique for analyzing the variable lengths of DNA fragments that result
from digesting a DNA sample with a special kind of enzyme. This enzyme, a
restriction endonuclease, cuts DNA at a specific sequence pattern know as a
restriction endonuclease recognition site. The presence or absence of certain
recognition sites in a DNA sample generates variable lengths of DNA fragments,
which are separated using gel electrophoresis. They are then hybridized with DNA
probes that bind to a complementary DNA sequence in the sample.
RFLP is one of the original applications of DNA analysis to forensic investigation.
With the development of newer, more efficient DNA-analysis techniques, RFLP is not
used as much as it once was because it requires relatively large amounts of DNA. In
addition, samples degraded by environmental factors, such as dirt or mold, do not
work well with RFLP.
PCR Analysis
PCR (polymerase chain reaction) is used to make millions of exact copies of DNA
from a biological sample. DNA amplification with PCR allows DNA analysis on
biological samples as small as a few skin cells. With RFLP, DNA samples would
have to be about the size of a quarter. The ability of PCR to amplify such tiny
quantities of DNA enables even highly degraded samples to be analyzed. Great
care, however, must be taken to prevent contamination with other biological
materials during the identifying, collecting, and preserving of a sample.
STR Analysis
Short tandem repeat (STR) technology is used to evaluate specific regions (loci)
within nuclear DNA. Variability in STR regions can be used to distinguish one DNA
profile from another. The Federal Bureau of Investigation (FBI) uses a standard set
of 13 specific STR regions for CODIS. CODIS is a software program that operates
local, state, and national databases of DNA profiles from convicted offenders,
unsolved crime scene evidence, and missing persons. The odds that two individuals
will have the same 13-loci DNA profile is about one in one billion.
Mitochondrial DNA Analysis
Mitochondrial DNA analysis (mtDNA) can be used to examine the DNA from
samples that cannot be analyzed by RFLP or STR. Nuclear DNA must be
extracted from samples for use in RFLP, PCR, and STR; however, mtDNA
analysis uses DNA extracted from another cellular organelle called a
mitochondrion. While older biological samples that lack nucleated cellular
material, such as hair, bones, and teeth, cannot be analyzed with STR and
RFLP, they can be analyzed with mtDNA. In the investigation of cases that
have gone unsolved for many years, mtDNA is extremely valuable.
All mothers have the same mitochondrial DNA as their daughters. This is
because the mitochondria of each new embryo comes from the mother's
egg cell. The father's sperm contributes only nuclear DNA. Comparing the
mtDNA profile of unidentified remains with the profile of a potential maternal
relative can be an important technique in missing person investigations.
Y-Chromosome Analysis
The Y chromosome is passed directly from father to son, so the analysis of
genetic markers on the Y chromosome is especially useful for tracing
relationships among males or for analyzing biological evidence involving
multiple male contributors.
PCR
The Polymerase Chain Reaction
Let’s Take a Break
PCR
The Polymerase Chain Reaction
Repetition of this cycle will cause repeated replication of the target
THERE ARE MILLIONS OF DIFFERENT GENES OR
SEQUENCES WITHIN ANY DNA SAMPLE (BLOOD,
TISSUE, PLANT, ETC.).
A SPECIFIC SEQUENCE IS SELECTED TO BE
AMPLIFIED (RED ABOVE). THIS SEQUENCE CAN BE
ANY GENE OF INTEREST OR A NON-CODING
MARKER REGION OF DNA.
IN ORDER TO COPY THE SEQUENCE OR GENE, A
SHORT SEQUENCE ON EITHER SIDE OF THE
SECTION MUST BE KNOWN. THIS REGION (BLUE
ABOVE) WILL SERVE AS A PRIMER ATTACHMENT
SITE TO COPY THE DNA TARGET SEGMENT.
IN ORDER TO AMPLIFY A SPECIFIC FRAGMENT OF DNA,
SEVERAL THINGS ARE NEEDED, INCLUDING PRIMERS
AND DNA POLYMERASE,
AN ENZYME WHICH COPIES DNA. PRIMERS ARE SHORT
PIECES OF DNA OR RNA DESIGNED TO PAIR WITH
GENOMIC DNA AT A SPECIFIC ATTACHMENT SITE FOR
THE MAIN PURPOSE OF HELPING THE DNA POLYMERASE
BIND AT THE DESIRED SECTION.
WITHOUT A SHORT PIECE OF DNA(OR
RNA) TO ATTACH TO, DNA POLYMERASE
CAN NOT COPY A DNA STRAND.
NUCLEOSIDE TRIPHOSPHATES, THE BUILDING BLOCKS OF
DNA ARE ALSO NEEDED.
EACH NUCLEOSIDE TRIPHOSPHATE CONSISTS OF:
A BASE (ADENINE, THYMINE, CYTOSINE OR
GUANINE).
A SUGAR AND THREE PHOSPHATES.
PCR REQUIRES SEVERAL CYCLES OF AMPLIFICATION. EACH CYCLE CONSISTS OF
THREE TEMPERATURE CHANGES.
THE STARTING TEMPERATURE (95 C) SEPARATES THE DNA STRANDS.
A LOWERED TEMPERATURE (50-60 C) ALLOWS PRIMERS TO BIND TO
COMPLEMENTARY SEQUENCES IN THE DNA.
A SLIGHTLY HIGHER TEMPERATURE (72 C) ALLOWS DNA POLYMERASE
TO ATTACH TO THE PRIMERS AND COPY THE DNA STRANDS
(EXTENSION).
DNA STRANDS ARE SEPARATED BY HEATING @ 94o C.
THE TEMPERATURE IS LOWERED TO 54oC TO
ALLOW PRIMERS TO PAIR WITH
COMPLEMENTARY DNA SEQUENCES.
MAKING NEW DNA MOLECULES:
DNA POLYMERASE ATTACHES TO THE
PRIMERS @ 72 C.
DNA POLYMERASE ADDS NUCLEOSIDE
TRIPHOSPHATES TO THE PRIMERS TO COPY THE
DNA STRANDS.
COPYING IS COMPLETED FOR EACH STRAND.
THE PROCESS IS REPEATED IN THE NEXT CYCLE.
THE TEMPERATURE IS RAISED AGAIN TO SEPARATE
THE DNA STRANDS.
THE TEMPERATURE IS LOWERED TO ALLOW PRIMERS
TO ANNEAL. DNA POLYMERASE ATTACHES TO
THE PRIMERS AND DNA IS COPIED TO MAKE
4 STRANDS OF DNA.
STRs Are Used in Identity Testing
“Short
Tandem
Repeat
sequence”
...atatatacaacttactaccatata
ccgattacgatcgaattataccgcgga
cgtagtaatgacgatgaagtaactata
tatatatatatatatatatatatatatatata
tatatatatatatatatatatatatatatata
tatatatatatatatatatatatatatatata
tatatatatatatatatatatatatatatata
tatatatatatatatatatatatatatatata
tatatatatatatatatatatatatatat
atactacctaccagggaggagata...
CODIS 13 Core STR Loci with Human
Chromosomal Positions
TPOX
D3S1358
D8S1179
D5S818
FGA
CSF1PO
TH01
VWA
D7S820
AMEL
D13S317
D16S539
D18S51
D21S11
AMEL
THE PROCESS OF COPYING DNA STRANDS IS REPEATED
32-35 TIMES. WITH EACH AMPLIFICATION CYCLE,
THE NUMBER OF COPIES OF THE DNA SEQUENCE IS
DOUBLED UNTIL MILLIONS OF COPIES HAVE BEEN MADE.
Over 1 million copies are generated in
32 cycles of this chain reaction
These copies can easily be detected
by gel electrophoresis. The size of the
DNA fragement should be that of
thetarget sequence
What is CODIS? Combined DNA Index System
CODIS is a computer software program that operates local, State, and national
databases of DNA profiles from convicted offenders, unsolved crime scene
evidence, and missing persons. Every State in the Nation has a statutory provision
for the establishment of a DNA database that allows for the collection of DNA
profiles from offenders convicted of particular crimes. CODIS software enables
State, local, and national law enforcement crime laboratories to compare DNA
profiles electronically, thereby linking serial crimes to each other and identifying
suspects by matching DNA profiles from crime scenes with profiles from convicted
offenders. The success of CODIS is demonstrated by the thousands of matches that
have linked serial cases to each other and cases that have been solved by matching
crime scene evidence to known convicted offenders.
The missing persons index consists of the unidentified persons index and the
reference index. The unidentified persons index contains DNA profiles from
recovered remains, such as bone, teeth, or hair. The reference index contains DNA
profiles from related individuals of missing persons so that they can be periodically
compared to the unidentified persons index. All samples for this index are typed
using mtDNA and STR DNA analysis (if possible) to maximize the power of
advancing technology.
PCR Copies DNA Exponentially
through Multiple Thermal Cycles
Original DNA target region
Thermal cycle
In 32 cycles at 100% efficiency, 1.07 billion
copies of targeted DNA region are created
Laboratory PCR Instrument
INPUT:
Sample DNA, PCR enzymes,
primers, individual nucleotide
building blocks (and maybe
fluorescent labels)
OUTPUT:
Specific DNA fragments amplified
millions of times for easy visualization
With sizes that vary between individuals
Multiplex PCR
• Over 10 Markers Can Be
Copied at Once
• Sensitivities to levels less than
1 ng of DNA
• Ability to Handle Mixtures
and Degraded Samples
• Different Fluorescent Dyes
Used to Distinguish STR
Alleles with Overlapping Size
Ranges
Available Kits for STR Analysis
• Kits make it easy for labs to just add DNA
samples to a pre-made mix
• 13 CODIS core loci
– Profiler Plus and COfiler (PE Applied Biosystems)
– PowerPlex 1.1 and 2.1 (Promega Corporation)
• Increased power of discrimination
– CTT (1994): 1 in 410
– SGM Plus™ (1999): 1 in 3 trillion
– PowerPlex ™ 16 (2000): 1 in 2 x 1017
Identity Testing Using PCR
Analysis of four different sections of the DNA
Possible conclusions:
S = size standards
V = victim’s DNA
1 = suspect #1 blood
2 = suspect #2 blood
3 = suspect #3 blood
E = evidence #1
S = size standards
A. Suspect 1 DNA was
at the scene
B. Suspect 2 DNA was
at the scene
C. Suspect 3 DNA was
at the scene
D. None were at the scene
E. Multiple suspects
were at the scene
F. Data are inconclusive
S
S = size standards
V = victim’s DNA
1 = suspect #1 blood
2 = suspect #2 blood
3 = suspect #3 blood
E = evidence #1
S = size standards
V
1
2
3
E
S
450
400
350
300
250
200
150
100
50
A complete match!
Case Study:
State v. Michael DeCorso
Homicide (no DNA)
Rape: DNA in semen samples from two
teenage female victims
DNA Profiles, Marker D10S28
C V D E1 E2 E3
C
C = Control
V = Victim
D = Defendant
E = Evidentiary sample
Population frequency of
defendant’s genotype =
1/50
DNA Profiles, Marker D4S139
C V D E1 E2 E3
C
C = Control
V = Victim
D = Defendant
E = Evidentiary sample
Population frequency of
defendant’s genotype =
1/90
DNA Profiles, Marker D5S110
C V D E1 E2 E3
C
C = Control
V = Victim
D = Defendant
E = Evidentiary sample
Population frequency of
defendant’s genotype =
1/10
DNA Profiles, Marker TH01
C V D E1 E2 E3
C
C = Control
V = Victim
D = Defendant
E = Evidentiary sample
Population frequency of
defendant’s genotype =
1/70
The information from each gel can
be combined to tell us how common
the DNA profile is in the general
population
1/50 x 1/90 x 1/10 x 1/70 = 1/3,150,000
Random match probabilities
Year Case
No. of
loci
Match
probability
1996
State v. DeCorso
4 RFLPs
1/3,000,000
1997
State v. Higgins
5 RFLPs
1/400,000,000
1999
State v. Butterfield
9 STRs
1/215,000,000,000
1999
State v. Troyer
9 STRs
1/200,000,000,000
CODIS with 13 Markers- Probability of an identical match greater
than all the people who have ever been born in the history of the
earth
Forensic applications of
DNA based technologies
Fingerprinting
OJ Simpson
•
Identification
•
Paternity
•
Crime Solving
•
World wide
data base
•
Dramatic Growth In DNA-Based Forensics
Doesn't Translate Into Very Many Job
Opportunities
One set of
22 autosomes
(plus X)
One set of
22 autosomes
(plus X or Y)
Paternity Testing
Three children: The father claims he is not the father of the third child
Note: There are two alleles* for each
genetic marker
USE OF NON-HUMAN DNA
PERFECT DNA MATCH
WITH CAT STRS
(NY TIMES INTERNATIONAL
APRIL 24, 1997)
1994 HOMICIDE
PLASTIC BAG FOUND IN SHALLOW GRAVE WITH BLOODY
JACKET AND TRACE HAIRS
HAIRS BELONGED TO SUSPECT’S CAT, SNOWBALL
DOG DNA CONTRIBUTES
TO MURDER CONVICTIONS
SEATTLE, 1996 DOUBLE MURDER, 2 SUSPECTS
COUPLE TORTURED AND SHOT ALONG WITH PET DOG
DID BLOOD ON SUSPECT’S CLOTHING MATCH THE DOG?
1 IN 3 BILLION MATCH PROBABILITY IN RANDOM CANINE
POPULATION.
FORENSIC PLANT DNA
DNA MARKERS FOR PLANTS CAN BE USED TO LINK
EVIDENCE TO A CRIME SCENE
GRASS STAINS LINKED TO LAWNS
VEGTABLE DNA LINKED TO RESTURANTES
RARE OR UNUSUAL PLANTS LINKED TO VEHICLES
MARAJUANA
The CT CODIS Database collects two types of samples; (1)
Convicted Offender Samples that include all Felony Convictions
(since 03/01/04) and, (2) Forensic Unknowns that include any DNA
profile from an evidentiary sample that does not match the victim
or an elimination known. There are currently 10,793 offenders in
CT Database and over 1500 offender samples are added per
month. Currently there are how many felons on the CT database?
(A) 1 out of 50 males in CT
(B) 1 out of 500 males in CT
(B) 1 out of 1000 males in CT
(C) 1 out of 10,000 males in CT
(E) None of these answers is correct
25
20
15
10
5
0
1
2
3
4
5
CASE STUDY
Renee Pellegrino
Renee Pellegrino
• Renee Pellegrino’s found June 25,
1997,murder victim.
• Renee Pellegrino was an accomplished scholar
with a law degree, had become addicted to
crack cocaine and turned to prostitution. She
was 40 years old and pregnant when her naked
body was discovered in a cul-de-sac off
Waterford Parkway South, CT.
Renee Pellegrino
She had just been released from a three-week
prison stay when she was apparently picked up
by her killer in downtown New London on June
25, 1997, murdered and left naked on Parkway
South.
She had been strangled, and the killer had left her
body in what the judge described as an
"extreme" manner.
Renee Pellegrino
Cold Case
Dickie Anderson 2012
Charged
Police make arrest in Pellegrino cold case
June 1, 2010
13 years later
• Police charged Dickie E. Anderson Jr., 40, with murder in the
41-year-old Pellegrino's death.
• The arrest was the result of a cold-case investigation into
Pellegrino's death, which occurred on a dead-end street in
Waterford. The state Office of the Chief Medical Examiner ruled
that Pellegrino had been strangled.
• Anderson, who was 27 at the time of the crime, was arrested as
a result of an investigation conducted by the Southeastern
Connecticut Cold Case Unit. The arrest warrant has been
sealed.
Dickie Anderson 2012
Charged
Police make arrest in Pellegrino cold case
June 1, 2010
Dickie Anderson 2012
Charged
Police make arrest in Pellegrino cold case
June 1, 2010
Police used a combination of evidence to build the case
against the man accused in the 1997 murder of Renee
Pellegrino, including DNA, inconsistent statements
Dickie E. Anderson Jr. made to police over the years
and his own admission that he was with Pellegrino
shortly before her body was discovered in Waterford.
Dickie Anderson 2012
• Over 13 years a case was built using
DNA, inconsistent statements Anderson
had made and his admission that he was
with Pellegrino shortly before her body
was discovered in a cul-de-sac off
Parkway South, CT.
Dickie Anderson 2012
• This is not a trick question
• “If you had to vote right now in this case,
guilty or not guilty, what would you do?"
A. Guilty
B. Not Guilty
C. Can’t vote; I don’t know the facts of the case
Dickie Anderson 2012
• This is not a trick question
• “If you had to vote right now in this case,
guilty or not guilty, what would you do?"
This is a common question during jury selection, and many people are tempted
to respond, incorrectly, that they can't vote because they don't know the facts of
the case.
The correct answer, as supplied by a woman in New London CT who was
eventually selected to serve on the panel, is "not guilty."
Dickie Anderson 2012
In a second case against, police charged
Anderson with killing 29-year-old Michelle Comeau of
Norwich in 1998. Anderson was previously charged
with the murder of Renee Pellegrino. The Comeau
case did not involve DNA.
Both of the women had been working as prostitutes and
were victims of strangulation. Police found Comeau's
body dumped along an access road to the Norwich
Industrial Park near Dodd Stadium in May 1998.
Anderson has acknowledged he knew both victims.
He told police he had been with Pellegrino on the night
she disappeared from downtown New London.
Dickie Anderson 2012
• . Pellegrino had been strangled, and her killer had
posed her naked body. In May 1998, police found
Comeau's body dumped along an access road to the
Norwich Industrial Park near the Norwich-Franklin
town line. She too had been strangled. The state
claims her killer was in the process of posing her
naked body in a similar manner but was interrupted
and left the scene. Anderson's DNA was found on
Pellegrino's body, but there is no DNA evidence to link
him to the Comeau case. He knew both women and
admitted to having sex with them on the day they were
killed
Dickie Anderson 2012
• One inmate reported that Anderson admitted to killing
"Renee" and said several times that he would have
never done it if he had known Pellegrino was
pregnant. She was 17 weeks pregnant when she died,
according to the state Office of the Chief Medical
Examiner.
• Police also spoke with former girlfriends of Anderson
who told them he was rough during sex. One woman
said Anderson threatened her and said he had gotten
away with killing somebody. The woman said
Anderson said he fought with a prostitute who kept
asking him for money, and that he hit and killed the girl
in Bates Woods in New London.
Dickie Anderson 2012
• The police also interviewed a girlfriend who broke up
with Anderson in 2000. She recalled that twice
Anderson choked her so hard he left red marks on her
neck, the warrant says. She turned over to police a
picture of her injuries that she said a friend had taken
• Another former girlfriend, whom Anderson was
convicted of strangling in 2008, said the two had
argued about her getting a job and that Anderson
threw her to the floor and began choking her. She said
if police did not break into the apartment and
physically remove Anderson from her, she thinks she
would have died.
Dickie Anderson 2012
• . Dickie Anderson Jr., a self-confessed "trick
artist" who told police he often traded crack
cocaine for sex with prostitutes, was connected
by witnesses to both of the women he is
accused of killing.
• In 2008, the state forensic laboratory had notified
police that DNA taken from Pellegrino's body matched
a DNA sample that had been taken from Anderson.
The laboratory also found DNA on Pellegrino from an
unknown source.
Dickie Anderson 2012
• Anderson's previous convictions include:
• • Jan. 2007, third-degree assault
• • November 2005, violation of a protective order
• • September 2005, second-degree failure to appear in
court, second-degree threatening, second-degree
criminal mischief
• • May 2003, violation of probation, evading responsibility,
second-degree failure to appear in court, third-degree
assault
• • July 2002, interfering with a police officer
• • March 1999, second-degree assault
Dickie Anderson 2012
•
Dickie Anderson 2012
• This is not a trick question
• “If you had to vote right now in this case,
guilty or not guilty, what would you do?"
A. Guilty
B. Not Guilty
C. Can’t vote; I don’t know the facts of the case
Dickie Anderson 2012
•
Dickie Anderson 2012
• . Anderson convicted of one of two murders
•
• Jury decides he killed Pellegrino in Waterford cold case;
mistrial declared in deadlock over second prostitute
murder
The Death Penalty
The Death Penalty
A. For
B. Against
The Death Penalty
Capital punishment in Rhode Island
The Hollow Men
T. S. Eliot
Mistah Kurtz—he dead. A penny for the Old Guy
I.
We are the hollow men
We are the stuffed men
Leaning together
Headpiece filled with straw. Alas!
Our dried voices, when
We whisper together
Are quiet and meaningless
As wind in dry grass
Or rats’ feet over broken glass
In our dry cellar
Shape without form, shade without color,
Paralyzed force, gesture without motion;
Those who have crossed
With direct eyes, to death’s other Kingdom
Remember us—if at all—not as lost
Violent souls, but only
As the hollow men The stuffed men.
Can We Please Take a Break?