Forensic Genetics and Molecular Basis of
Application of DNA Finger-Printing to
Forensic Science as New Tools to Solve
Criminal Dilemmas
Clinical Biochemistry, Faculty of
Medicine, University of Tabuk, KSA.
At the end of the lecture
New aspects of identification used in forensic
practice
DNA fingerprints and its role in human
identification
Usage of DNA fingerprints in cases of identification
in rape and disputed paternity cases.
Medico-legal importance of the usage of DNA
fingerprinting as a new tool in forensic practice.
Genetic fingerprinting
The pattern of bands in a gel electrophoresis is known as a
genetic fingerprint or a ‘genetic profile’
DNA analysis can be used for catching criminals, establishing
parentage, finding how closely organisms are related and many other
applications.
If a genetic fingerprint found in a sample of blood or other tissue
at the scene of a crime matches the genetic fingerprint of a suspect,
this can be used as evidence
A DNA sample can be obtained from the suspect using blood, cheek
epithelial cells taken from the mouth lining or even the cells clinging
to the root of a hair
Early 1980s: Restriction Fragment Length
Polymorphism (RFLP)
Genetic variation in the distance
between restriction enzyme sites
Template DNA digested by enzymes,
electrophoresed, detected via
Southern blotting
Power of discrimination in the range of
106-108 for a six probe analysis
Mechanisms
for RFLPs
12
Appearance of separated fragments on gel
These bands will
contain the shorter
DNA fragments
These bands will
contain the longer
DNA fragments
Prof.E.J.Wood
©©Prof. E. Wood
starting positions Appearance of bands
Disadvantage
RFLP testing requires a relatively
large amount of HMW DNA (~50ng =
thousands of cells)
Not ideal for forensic evidence, in
which small, degraded samples are
common
PCR To The Rescue!
Polymerase Chain Reaction =
molecular Xeroxing
Three temperature phases, carried out
in a Thermal Cycler, replicate or
“amplify” the desired DNA fragment(s)
Applications
Diagnosing Disease
Applications
Paternity Testing
20
Genetic fingerprint of …
1 mother
2 child
3 possible father A
4 possible father B
There is a match between one of
the child’s restriction fragments
and one of the mother’s.
There is also a match between the
child’s other fragment and one from
possible father A.
Neither of the child’s restriction
1 2 3 4 fragments match those of possible
father B
Starting position of sample Paternity test
The Current Method of Choice: Autosomal
Short Tandem Repeats
Non-coding, tetranucleotide sequences
which vary greatly from person to
person in the number of repeating units
Requires <1ng of DNA to type 13-15
STR loci
Power of discrimination ranges from
1014-1023. World population is 109 so
bring on the database!
Applications
Forensics
Applications
Victim
DNA
Crime Fingerprinting
Scene Animation
Suspect
STR data (cont’d)
STR TYPING SUMMARY SHEET
Date: DNA Analyst / Serial #: DR #:
9/24/1999 MATTHIES V9780 00-00-00001
Item # AMEL D3S1358 vWA FGA D8S1179 D21S11 D18S51 D5S818 D13S317 D7S820 D16S539 THO1 TPOX
X, Y 17 8, 10
25(S) 15, 17 23, 26 14, 15 26 12, 15 10 9, 13 9, 10 8, 9 9, 10
X, Y 17 8, 10
X 15, 17 28, 11
25(E) 16, 18 19, 26 15 14, 16 8, 13 12 11, 12 7, 8 11
X 15, 17 32.2 11
X 15, 17 28, 11
VICTIM 16, 18 19, 26 15 14, 16 8, 13 12 11, 12 7, 8 11
X 15, 17 32.2 11
X, Y 17 8, 10
SUSPECT 15, 17 23, 26 14, 15 26 12, 15 10 9, 13 9, 10 8, 9 9, 10
X, Y 17 8, 10
“The DNA profile obtained from Item 25(S) matches the
DNA profile of the suspect. The combination of genetic
marker types exhibited by Item 25(S) and the suspect
occurs in approximately one in one hundred quadrillion
(1017) individuals…”
How are these astronomical figures derived?
The product rule: combined probability of a series of independent
events is determined by multiplying the probabilities of each event.
STR loci are inherited independently (unlinked)
Homozygous loci: p2 (same allele inherited from mother and father)
Heterozygous loci: 2pq (either allele could be inherited from either
parent)
p(17)2 x 2p(15)q(17) x 2p(23)q(26)….
(.223)2 x 2(.083)(.25) x 2(.14)(.02) = .000013, which is equivalent
to a probability of one in 76,000 using just 3 of the 13 loci!
Applications
Genealogy
Mitochondrial DNA (mtDNA)
Pros
Single-cell sensitivity because each cell contains
~1000 mitochondria
Especially useful for shed hairs, burnt remains
Can be used to establish kinship directly because
entire complement of mtDNA is maternally inherited
Mitochondrial DNA (mtDNA)
The Combined DNA Index System (CoDIS)
A database of DNA profiles from violent
felons and crime scene samples
Laws concerning who is eligible for the
database vary from state to state
Database currently contains about
2,038,470 felons and 93,956 crime scene
profiles (19,00 hits so far)
Advantage of CoDIS
Extremely powerful investigative
tool, linking crimes, and pulling
suspects out of thin air!
Can prevent, as well as solve
crimes!
Disadvantage of CoDIS
(What the FBI doesn’t want you to know.)
DNA mixtures and degraded DNA profiles have
lead to spurious matches
Stringent laws explicitly permit databasing
innocent people
Adding arrestees to database violates presumption
of innocence
However, the prosecution rate on case to offender
matches is shockingly low! (~10%)
“Specialized” PCR-based systems
mtDNA
Y-STRs
SNPs
Y-STRs
Problem:
~99% of violent crimes are committed by men
DNA Mixtures of male suspect and female victim can
pose an analytical challenge, especially when the
female contribution is much greater than the male =
preferential amplification
Solution:
Test for markers found only on the Y-chromosome.
Only male DNA is amplified!
Single Nucleotide Polymorphisms (SNPs)
Point mutations (base substitutions) found in 1% or
more of the population
1.8 million identified in human genome
Detected on micro-array plates with fluorescent tags
(all or nothing response)
~50 SNPs provides same power of discrimination as
13 STR loci
Certain SNPs used as predictors of
ancestry/ethnicity by a private sector lab (DNA
Witness)
SNP Flaws
Privacy issues - unlike STRs, SNPs can be
correlated with susceptibility/resistance to
diseases
Requires a relatively large quantity of DNA for
robust assay
