DNA PROFILING
Principles, Mechanism, Applications and Forensic Significance
An Academic Assignment
Submitted as part of coursework in Forensic Science / Molecular Biology
Department of Biotechnology / Forensic Science
Academic Year 2025–2026
Table of Contents
1. Introduction
2. Historical Background
3. Basic Principle
4. Mechanism / Working Procedure
5. STR Loci and CODIS
6. Applications
7. Advantages
8. Disadvantages / Limitations
9. Illustrative Example
10. Conclusion
11. References
DNA Profiling — Assignment
1. Introduction
DNA profiling, also known as DNA fingerprinting or genetic profiling, is a scientific technique used to identify
individuals based on unique patterns present in their DNA. Although more than 99.9% of human DNA sequence is
identical between any two people, certain regions of the genome are highly variable, and these regions form the
basis of DNA profiling. Except in the case of identical (monozygotic) twins, no two individuals share the same
complete DNA profile, making the technique an exceptionally powerful tool for personal identification.
Today, DNA profiling is central to forensic science, paternity testing, disaster victim identification, and the
exoneration of wrongly convicted individuals, and it represents one of the most significant contributions of
molecular biology to criminal justice systems worldwide.
2. Historical Background
DNA profiling was developed in 1984 by Sir Alec Jeffreys at the University of Leicester, who discovered that
certain repetitive DNA sequences varied significantly between individuals. Using a method called Restriction
Fragment Length Polymorphism (RFLP), Jeffreys demonstrated that these variable regions could serve as a unique
genetic identifier. The technique was first applied forensically in 1986 in the Enderby murders case in the United
Kingdom, where it both convicted the true perpetrator and exonerated a wrongly suspected individual — a landmark
moment establishing DNA evidence in criminal investigation. Since then, the field has progressed from RFLP to
faster, more sensitive PCR-based Short Tandem Repeat (STR) analysis, which remains the standard approach used
today.
3. Basic Principle
DNA profiling exploits highly polymorphic, non-coding regions of the genome known as Short Tandem Repeats
(STRs) — short DNA sequences of two to six base pairs that repeat a variable number of times at specific
chromosomal locations (loci). The number of repeat units at a given locus differs from person to person, and by
examining multiple STR loci simultaneously, the probability that two unrelated individuals share an identical profile
becomes vanishingly small — often less than one in several billion.
Figure 1: Concept of a Short Tandem Repeat (STR) locus — two individuals with a different number of repeat units at the same
locus.
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DNA Profiling — Assignment
4. Mechanism / Working Procedure
A typical DNA profiling analysis proceeds through the following defined steps, from sample collection to final
statistical interpretation:
• Sample Collection: biological material is collected from a crime scene, a suspect, or a reference source —
blood, saliva, semen, hair follicles, skin cells, or bone.
• DNA Extraction: DNA is isolated from the cellular material using organic (phenol-chloroform), Chelex, or
silica-column-based extraction methods.
• Quantification: the concentration and quality of extracted DNA is measured to ensure sufficient template for
amplification.
• PCR Amplification: the target STR loci are exponentially amplified using the Polymerase Chain Reaction with
fluorescently labelled primers that flank each repeat region.
• Capillary Electrophoresis: the amplified, fluorescently tagged fragments are separated by size in a capillary
electrophoresis instrument, with smaller fragments migrating faster than larger ones.
• Allele Calling: a laser detector records the fluorescent signal as each fragment passes the detection window,
producing an electropherogram; software compares peak positions to a reference allelic ladder to assign
specific repeat-number allele calls.
• Profile Comparison and Statistical Analysis: the resulting profile is compared against a reference sample or
database entry, and a Random Match Probability (RMP) is calculated to express the statistical significance of a
match.
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DNA Profiling — Assignment
Figure 2: Stepwise workflow of a DNA profiling analysis, from sample collection to reporting.
4.1 Representative Electropherogram
The final readout of a capillary electrophoresis-based STR analysis is an electropherogram — a plot of fluorescence
intensity against fragment size, in which each peak corresponds to a specific allele at a given locus. A single peak at
a locus indicates homozygosity (identical alleles inherited from both parents), while two peaks indicate
heterozygosity (different alleles inherited from each parent).
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DNA Profiling — Assignment
Figure 3: Representative electropherogram showing allele peaks at three STR loci (D3S1358, TH01, D21S11).
5. STR Loci and CODIS
Forensic laboratories analyze a standardized panel of STR loci to ensure that profiles generated in different
laboratories, and even different countries, remain comparable. In the United States, the FBI's Combined DNA Index
System (CODIS) uses a core set of 20 STR loci (expanded from an original 13 in 2017), together with the
Amelogenin marker used to determine biological sex. Similar standardized panels are used by national DNA
databases in other countries, such as the UK's National DNA Database (NDNAD).
Locus Chromosome Notes
D3S1358 Chromosome 3 Core CODIS locus
TH01 Chromosome 11 One of the first STR loci used
D21S11 Chromosome 21 Highly polymorphic core locus
vWA Chromosome 12 Core CODIS locus
Used for sex determination, not
Amelogenin X/Y chromosomes
identity
6. Applications
• Forensic investigation — matching crime scene evidence (blood, semen, hair, saliva) to suspects or victims
• Criminal database screening — comparing profiles against national databases such as CODIS (USA) or
NDNAD (UK) to identify repeat offenders or solve cold cases
• Paternity and kinship testing — establishing biological relationships in legal, immigration, or inheritance
disputes
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DNA Profiling — Assignment
• Missing persons and disaster victim identification — identifying remains in mass disasters by comparison with
relatives' DNA
• Exoneration of the wrongly convicted — post-conviction DNA testing has overturned hundreds of wrongful
convictions worldwide
• Immigration and family reunification — proving genetic relationships for visa or immigration claims
• Wildlife forensics and conservation — identifying species or individual animals in poaching investigations
• Anthropology and ancestry studies — tracing human migration patterns using ancient and modern DNA
7. Advantages
• Extremely high discriminatory power, with random match probabilities often below one in several billion
• Requires only minute biological samples, such as a single hair root or a few skin cells
• Highly standardized and reproducible across laboratories through common STR loci panels
• Rapid turnaround using automated PCR and capillary electrophoresis systems
• Effective even with partially degraded samples, owing to the amplifying power of PCR
• Provides objective, statistically quantifiable evidence admissible in courts of law
8. Disadvantages / Limitations
• Sample contamination or degradation can produce partial or unreliable profiles
• Cannot distinguish between identical (monozygotic) twins, who share essentially identical DNA
• Mixed samples from multiple contributors complicate interpretation and may require specialized statistical
models
• Requires a reference sample or database entry for meaningful comparison
• Risk of laboratory error or cross-contamination if strict quality-control procedures are not followed
• Raises ethical and privacy concerns regarding DNA database retention and potential misuse
• Low-quantity or degraded "touch DNA" samples increase the risk of allele drop-out or stutter artifacts,
requiring careful expert interpretation
9. Illustrative Example
In a criminal investigation, DNA extracted from blood found at a crime scene is amplified at the 20 CODIS STR
loci and separated by capillary electrophoresis. The resulting profile is compared against a database of previously
collected offender profiles. If, for example, 19 of the 20 loci match a specific database entry exactly, forensic
analysts calculate the random match probability and can report — with a very high degree of statistical confidence
— the likelihood that the DNA originated from that individual, providing evidence admissible in court.
10. Conclusion
DNA profiling has transformed forensic science, medicine, and biology by providing an exceptionally reliable
method for individual identification and relationship testing. Its foundation in highly polymorphic STR loci,
combined with the sensitivity of PCR amplification and the resolving power of capillary electrophoresis, allows
analysts to generate statistically robust genetic profiles from even trace amounts of biological material. Despite
certain limitations — including sample degradation, mixed-sample interpretation challenges, and ethical
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DNA Profiling — Assignment
considerations around DNA databases — DNA profiling remains one of the most powerful and widely trusted tools
in modern forensic and biomedical science.
11. References
1. Jeffreys, A. J., Wilson, V., & Thein, S. L. (1985). Individual-specific 'fingerprints' of human DNA. Nature,
316(6023), 76–79.
2. Butler, J. M. Forensic DNA Typing: Biology, Technology, and Genetics of STR Markers (2nd ed.). Academic
Press.
3. National Institute of Justice. DNA Evidence: Basics of Analyzing. U.S. Department of Justice.
4. Federal Bureau of Investigation. CODIS – Combined DNA Index System. FBI Laboratory Services.