DNA FINGERPRINTING

A POWERFUL FORENSIC TOOL FOR IDENTIFYING CRIMINIALS

Presented at
NEUROFRENZY - ENTENTE 2024

By Vinayak & Devarth

Origin & Introduction
DNA fingerprinting, also known as DNA profiling, was first developed by Sir Alec Jeffreys in 1984 at the University of Leicester, UK.

Discovery of VNTRs (Variable Number of Tandem Repeats):

Though around 99 percent of the Human genome is same, Variable Number of Tandem Repeats (VNTRs), which varied greatly between
individuals. This high degree of variability made it possible to distinguish between different people.
DNA fingerprinting is unique because it relies on the variability in the non-coding regions of DNA, where sequences are highly individual-specific.

The first practical use of DNA fingerprinting occurred in 1985, when it was used in an immigration case to confirm a family relationship in the UK. This Sir Alec John Jeffreys, is a British
non-criminal application demonstrated the potential of DNA fingerprinting for personal identification. geneticist known for developing
techniques for genetic fingerprinting
DNA fingerprinting was then used in a criminal investigation for the first time in 1986, in the case of two murders in Leicestershire, England. It
and DNA profiling which are now used
was instrumental in identifying the true perpetrator, Colin Pitchfork, and exonerating an innocent suspect​
worldwide in forensic science to assist
police detective work and to resolve
Application in Forensic Science: paternity and immigration disputes.
DNA fingerprinting quickly gained recognition as a powerful forensic tool, helping solve numerous cases by identifying individuals from biological award ed v a rio us prizes in
He has been including
material left at crime scenes (blood, hair, semen, etc.). ute r Sc ie nc e,
the field of Comp
It has since become a standard method in forensic investigations, paternity testing, and genetic research. the Turing Award

The development of PCR (Polymerase Chain Reaction) in the 1990s enhanced the technique, allowing for the amplification of small or degraded DNA
samples and expanding the applicability of DNA fingerprinting in forensic investigations​.
 The patterns of STRs are analyzed to create a DNA profile. Each person will have a unique
Sample Collection
Biological samples like blood, saliva,
pattern or "fingerprint" because the number of repeats in the STR regions varies among
individuals.
Steps & Processes
hair, semen, or skin cells are
collected from a crime scene,
individual, or any source of
investigation. After electrophoresis, DNA fragments The DNA profile obtained from the sample is compared with other profiles. For forensic cases,
are passed through a gel or a capillary the profile may be compared to a suspect’s DNA or entered into databases like the Combined
DNA Index System (CODIS), where profiles from unsolved cases or known criminals are stored.
tube, and an electric field is applied.
A match between DNA profiles indicates a high probability that the DNA came from the same
Smaller DNA fragments move faster
individual​
DNA Extraction than larger ones, separating them based
DNA is extracted from the collected on size. The separated DNA fragments
cells using chemical treatments that are visualized using fluorescent dyes or
break down cell membranes and other labeling techniques.
isolate the DNA.
Flow
Chart
PCR Amplification (Polymerase Chain DNA Fragment Separation describing
Reaction) is used to amplify specific regions After amplification, the STR loci
of DNA, particularly Short Tandem Repeats (specific locations on the DNA) are the
(STRs). These are short repeating separated according to size using a
sequences found in the non-coding regions technique called gel electrophoresis or process
of DNA that vary greatly between capillary electrophoresis.
individuals.
 OLDER METHOD OF DNA FINGER PRINTING
INVOLVING SOUTHERN BLOTTING AND USE
OF RESTRICTION ENDO-NUCLEASE
Endonucleases are enzymes that
cleave the phosphodiester bonds
within DNA or RNA strands at specific
internal sites. Unlike exonucleases,
which trim nucleotides from the ends,
endonucleases cut at internal
positions, creating DNA or RNA
fragments.

image credits: infinity learn

SOUTHERN BLOTTING IS A LABORATORY TECHNIQUE USED TO DETECT SPECIFIC DNA SEQUENCES WITHIN A DNA SAMPLE. THE PROCESS INVOLVES:
1. DNA DIGESTION: DNA IS CUT INTO FRAGMENTS USING RESTRICTION ENZYMES.
2. GEL ELECTROPHORESIS: THE FRAGMENTS ARE SEPARATED BASED ON SIZE THROUGH AN AGAROSE GEL.
3. TRANSFER TO A MEMBRANE: THE SEPARATED DNA IS TRANSFERRED FROM THE GEL ONTO A NYLON OR NITROCELLULOSE MEMBRANE.
4. HYBRIDIZATION: A LABELED DNA PROBE COMPLEMENTARY TO THE TARGET SEQUENCE BINDS TO THE DNA OF INTEREST ON THE MEMBRANE.
5. DETECTION: THE BOUND PROBE IS VISUALIZED, REVEALING THE PRESENCE AND SIZE OF THE TARGET DNA.
 DNA FINGER PRINTING IN CONTEXT TO
THE CRIMINAL JUSTICE SYSTEM
1. Crime Scene Investigation
DNA collected from biological evidence at a crime scene (e.g., blood, saliva, hair) can be analyzed to create a DNA profile.
This profile can be compared to known suspects to establish a match or exclude individuals.

2. Suspect Identification
If a suspect's DNA matches the DNA profile obtained from evidence at the crime scene, it can provide strong evidence of their involvement in the
crime.
DNA evidence can link a suspect to the crime scene or victim, supporting charges against them.

3. Exoneration of Innocent Individuals

DNA fingerprinting can also be used to exonerate individuals wrongfully accused or convicted of a crime by comparing their DNA to the evidence
found at the scene.

4. Paternity and Family Relationship Testing

DNA fingerprinting can be used to establish familial relationships, which can help in criminal investigations involving family members or to clarify
potential suspects.

5. Database Matching
National DNA databases, such as the Combined DNA Index System (CODIS) in the U.S., store DNA profiles from convicted offenders and crime scenes.
DNA from new cases can be entered into these databases to check for matches with previously collected samples, helping to identify repeat
offenders.

6. Cold Cases
DNA fingerprinting can be applied to unresolved cases, allowing law enforcement to revisit and analyze old evidence with modern techniques.
Advances in DNA analysis have led to the solving of many cold cases that were previously deemed unsolvable.
CASE: TOMMY LEE ANDREWS

BACKGROUND: IN 1986, A SERIES OF RAPES TOOK PLACE IN ORLANDO, FLORIDA. VICTIMS DESCRIBED A SIMILAR
PATTERN: A MAN BREAKING INTO THEIR HOMES AND ASSAULTING THEM. DESPITE MULTIPLE VICTIMS PROVIDING
Case Study One
DESCRIPTIONS, THERE WAS LITTLE PHYSICAL EVIDENCE TYING ANYONE DIRECTLY TO THE CRIMES. HOWEVER, LAW
ENFORCEMENT RECOVERED SEMEN SAMPLES FROM SEVERAL CRIME SCENES.

ROLE OF DNA FINGERPRINTING: THIS CASE MARKED THE FIRST TIME DNA FINGERPRINTING WAS USED IN THE UNITED
STATES TO OBTAIN A CONVICTION. LAW ENFORCEMENT WORKED WITH FORENSIC SCIENTISTS TO ANALYZE THE SEMEN
SAMPLES, WHICH WERE FOUND TO MATCH THE DNA OF TOMMY LEE ANDREWS, WHO HAD A PRIOR HISTORY OF
CRIMINAL ACTIVITY AND WAS UNDER INVESTIGATION.
USING DNA PROFILING, THE PROSECUTION SHOWED A DIRECT LINK BETWEEN THE DNA EVIDENCE FROM THE CRIME
SCENES AND ANDREWS’ DNA. THIS WAS REVOLUTIONARY BECAUSE, AT THE TIME, NO OTHER FORM OF PHYSICAL
EVIDENCE COULD SO DEFINITIVELY LINK A SUSPECT TO A CRIME.

OUTCOME: IN 1987, ANDREWS WAS CONVICTED OF MULTIPLE COUNTS OF RAPE, BURGLARY, AND OTHER CRIMES.

IMPACT:

FIRST U.S. CONVICTION USING DNA EVIDENCE: THIS CASE SET A LEGAL PRECEDENT IN THE UNITED STATES FOR THE
ADMISSIBILITY OF DNA EVIDENCE IN COURT.
THE SUCCESS OF THIS CASE HELPED ESTABLISH DNA FINGERPRINTING AS A RELIABLE AND POWERFUL FORENSIC
TOOL IN CRIMINAL INVESTIGATIONS.
AFTER THIS CASE, DNA EVIDENCE BECAME A STANDARD METHOD IN FORENSIC SCIENCE, LEADING TO THE
CONVICTION OF COUNTLESS VIOLENT CRIMINALS.
THE CASE OF VISHNU TIWARI (2000)

BACKGROUND: VISHNU TIWARI WAS SENTENCED TO LIFE IMPRISONMENT IN 2000

Case Study Two
FOR THE ALLEGED RAPE OF A WOMAN IN LALITPUR, UTTAR PRADESH. HE SPENT 20
YEARS BEHIND BARS, CONSISTENTLY CLAIMING HE WAS INNOCENT.

ROLE OF DNA FINGERPRINTING: IN 2021, THE ALLAHABAD HIGH COURT REVIEWED THE
CASE. DNA FINGERPRINTING, WHICH HAD NOT BEEN USED DURING HIS ORIGINAL TRIAL,
WAS APPLIED TO REEXAMINE THE EVIDENCE. THE DNA RESULTS FAILED TO
CORROBORATE THE ALLEGATIONS, AND SEVERAL DISCREPANCIES WERE FOUND IN THE
INVESTIGATION. BASED ON THESE FINDINGS, THE COURT DECLARED TIWARI INNOCENT
AND RELEASED HIM AFTER TWO DECADES OF WRONGFUL IMPRISONMENT.

IMPACT: THIS CASE HIGHLIGHTS THE GROWING RECOGNITION OF DNA FINGERPRINTING

IN INDIAN COURTS AS A TOOL TO CORRECT WRONGFUL CONVICTIONS AND ENSURE
JUSTICE FOR THOSE WRONGFULLY ACCUSED.
 advancement in DNA Fingerprinting
Southern Blotting Polymerase Chain Reaction (PCR)
Process: Process: Conclusion
a. DNA is digested with restriction enzymes to create a. DNA is denatured to separate strands.
fragments. b. Primers anneal to the target sequences. Higher Sensitivity: PCR can detect
b. The fragments are separated by gel electrophoresis. c. DNA polymerase synthesizes new strands, amplifying and amplify minute quantities of
c. DNA is transferred from the gel to a membrane the target region. DNA, making it effective for
(blotting). d. This cycle is repeated. analyzing samples with low DNA
d. A labeled DNA probe that is complementary to the Advantages:
content (e.g., forensic
target sequence is hybridized to the membrane. Sensitivity: Requires only a small amount of starting
evidence).
e. Detection is performed to visualize the presence of DNA (even a single copy can be amplified).
the target DNA. Faster Results: PCR can provide
Speed: The entire process can be completed in a few
Limitations: hours. results within hours, whereas
Requires relatively large amounts of DNA. Simplicity: Less labor-intensive than Southern blotting. Southern blotting can take days
Time-consuming and labor-intensive. Specificity: Can amplify specific regions of interest, due to multiple steps involved.
Limited sensitivity compared to PCR. even in complex mixtures of DNA. Ease of Use: PCR is more
straightforward and requires
fewer steps compared to the
multi-step process of Southern
blotting.
 advancement in DNA Fingerprinting
Slab Gel Electrophoresis Capillary Electrophoresis
Setup: In slab gel electrophoresis, a gel (typically agarose) is Setup: Capillary electrophoresis uses thin capillaries filled Conclusion
cast in a flat, rectangular mold, forming a slab. with a gel.
Process: Process: Resolution and Speed: Capillary
Samples are loaded into wells at one end of the gel. Samples are injected into the capillary. electrophoresis is generally
An electric current is applied, causing charged molecules An electric current causes the charged molecules to better due to its higher
to migrate through the gel. migrate through the capillary. resolution and faster analysis
The gel matrix creates a sieve effect, allowing smaller The smaller diameter of the capillary allows for high-
molecules to move faster than larger ones. time.
resolution separation.
Visualization: After electrophoresis, the gel is stained (e.g., with Automation and Efficiency:
Visualization: Detection occurs in real-time as the molecules
ethidium bromide for DNA) to visualize the separated bands. Capillary systems can be
exit the capillary, often using fluorescence or UV detection.
Limitations: Advantages: automated, making them more
Resolution can be lower for very small differences in size. Faster analysis time. efficient.
Requires longer run times and can have issues with heat Sample Size: If larger sample
Higher resolution due to the narrow capillary.
generation, which may affect the results. volumes are required, slab gel
More susceptible to contaminantes.
may be preferred, although it is
generally slower and less
sensitive.
 advancement in DNA Fingerprinting

Slab Gel electrophoresis Capillary electrophoresis

image credits: Byju’s image credits: Davidson College
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Next-Generation Sequencing (NGS)

Next-Generation Sequencing (NGS)

NGS, also known as massively parallel sequencing (MPS), represents a significant advancement in DNA sequencing technology. It enables
the rapid and simultaneous sequencing of millions of DNA fragments, making it a revolutionary tool in forensic science.

Advantages:
High Throughput: NGS can analyze a large number of samples simultaneously, making it cost-effective and efficient.
Sensitivity and Specificity: It provides greater sensitivity and specificity than traditional sequencing methods, which is crucial for
forensic analysis, especially when dealing with degraded or low-quality samples​.
Complex Mixture Analysis: NGS can effectively handle complex DNA mixtures, allowing for better interpretation of mixed samples

Methodology:
NGS typically involves a workflow that includes sample preparation, library construction, amplification (often via PCR),
sequencing, and data analysis.
It allows for the simultaneous amplification of multiple STR marker types in a single run, significantly speeding up the analysis​
Phenotypic profiling

Phenotypic profiling is a technique used to predict physical characteristics (phenotypes) based on genetic
information. This approach analyzes specific genetic markers associated with observable traits such as eye color,
hair color, skin tone, and even certain facial features.

Phenotypic traits are influenced by multiple genes, each contributing to the overall appearance of an individual.
By examining single nucleotide polymorphisms (SNPs) and other genetic variations, researchers can associate
specific genes with particular traits.
In criminal investigations, phenotypic profiling can help create a visual description of an unknown individual
based on their DNA. This can assist law enforcement in identifying suspects or victims when traditional
identification methods are unavailable.

Methodology:
DNA Extraction: DNA is extracted from biological samples such as blood, saliva, or hair.
Genotyping: Specific genetic markers associated with phenotypic traits are analyzed.
Bioinformatics tools are used to interpret the genetic data and predict phenotypic traits based on established
associations between genetic markers and observable characteristics.
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TEAM A1