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DNA FINGERPRINTING

Article in European Journal Pharmaceutical and Medical Research · July 2017

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 ejpmr, 2017,4(8), 244-246 SJIF Impact Factor 4.161
Review Article
Singla et al. EUROPEAN JOURNAL OF PHARMACEUTICAL
European Journal of Pharmaceutical and Medical Research
AND MEDICAL RESEARCH ISSN 2394-3211
www.ejpmr.com EJPMR

DNA FINGERPRINTING

Dr. Kamal Singla1*, Dr. Yashoda Rani2, Dr. Kuldeep Panchal1, Dr. Rishabh Kumar Singh3,
Dr. Vivek Chouksey3
1
Senior Resident, Deptt. of Forensic Medicine, Lady Hardinge Medical College, New Delhi.
2
Head of Deptt, Deptt. of Forensic Medicine, Lady Hardinge Medical College, New Delhi.
3
Junior Resident, Deptt. of Forensic Medicine, Lady Hardinge Medical College, New Delhi.

*Corresponding Author: Dr. Kamal Singla

Senior Resident, Deptt. of Forensic Medicine, Lady Hardinge Medical College, New Delhi.

Article Received on 10/06/2017 Article Revised on 30/06/2017 Article Accepted on 20/07/2017

ABSTRACT
DNA profiling (DNA fingerprinting) is a technique employed by forensic scientists to assist in the identification
of individuals by their respective DNA profiles. DNA profiling should not be confused with full genome
sequencing. Every individual in the world can be identified at the molecular level on the basis of an extremely high
level of polymorphism in the sequence of his or her DNA, which he or she inherits from his or her biological
parents and is identical in every cell of the body. Although 99.9% of human DNA sequences are the same in every
person, enough of the DNA is different that it is possible to distinguish one individual from another, unless they
are monozygotic twins. DNA profiling uses repetitive sequences that are highly variable called variable number
tandem repeats (VNTRs) in particular short tandem repeats (STRs). VNTRs loci are very similar between closely
related humans, but are so variable that unrelated individuals are extremely unlikely to have the same VNTRs. The
application of DNA profiling in the criminal justice system is an important issue in criminal investigators today.
The technology is changing rapidly and several new techniques are becoming available. DNA profiling has been
described as a powerful breakthrough in forensic science. The forensic use of DNA profiling is a major
contribution to a technology which can help not only in including the culprit but also to exclude the innocent.

KEYWORDS: DNA, STRs, VNTRs.

INTRODUCTION polymorphism[4] which is exploited in DNA profiling

The DNA profiling technique was first reported in process.
1986[1] by Sir Alec Jeffreys at the University of Leicester
in England, United Kingdom[2] and is now the basis of Sample can be of two types[5]
several national DNA databases. Dr. Jeffrey’s genetic 1. Fresh un-degraded sample- As in paternity testing
fingerprinting was made commercially available in 1987, when fresh blood sample from putative father, mother &
when a chemical company, Imperial Chemical Industries child is available.
(ICI), started a blood testing centre in the UK.[3] Human 2. Degraded sample- As in old dried blood or seminal
body contains 60 trillion cells (6 x 1013). Each human stains generally found at the crime scene.
diploid cell contains 23 pair of chromosomes (46) of
which half is derived from each parent. Total DNA in a Conditions Causing DNA Degradation[5]
human haploid cell is 3 x 109 base pairs. About 99.9% of DNA breaks into smaller fragments when it is exposed to
DNA code is the same for all humans. It is only rest biological contaminants, chemicals, dirt, fungus, heat,
0.1% which is the basis of DNA profiling and all of this light (sunlight or UV rays) and moisture. Their average
lies in the non-coding region. Non coding DNA (junk size may become smaller than the area of interest (allele
DNA) constitutes 97% of the total nuclear DNA while length at a particular locus). If the average size of DNA
rest is coding DNA (3%).[4] fragments in a degraded sample is smaller than the allele
length, the allele would not be detected.
The substantial amount of non-coding DNA shows a
peculiarity. It consists of large arrays (alleles) of tandem In degraded sample average size of DNA fragments is
repeats of nitrogenous bases. This repetitive DNA <500bp, thus in such a sample STR & VNTR are
constitutes approximately 50% of the human genome. detectable while satellite DNA may or may not. Thus
The DNA on either side of repeats is called flanking different techniques are used for fresh and degraded
DNA. The number of repeats is different in different samples. In fresh samples - Satellite DNA & VNTR are
person and this property is known as tandem repeat

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Singla et al. European Journal of Pharmaceutical and Medical Research

detected by RFLP while in degraded samples – Also, Karl Brown's original technique looked at many
microsatellites are detected by STR analysis using PCR. mini-satellite loci at the same time, increasing the
observed variability, but making it hard to discern
Classification of tandem repeats[6] individual alleles (and thereby precluding parental
1. STRs (Short tandem repeats or micro-satellites) testing). These early techniques have been supplanted by
If the repeating unit is of the length between 1-6bps PCR-based assays.
(base pairs) then the sequence is called short tandem
repeats. STRs analysis: The system of DNA profiling used today
is based on PCR and uses short tandem repeats (STRs).
2. VNTRs (Variable number tandem repeats or mini- This method uses highly polymorphic regions that have
satellites): If the repeating unit is of the length between short repeated sequences of DNA (the most common is 4
7-100bps (base pairs) then the sequence is called variable bases repeated, but there are other lengths in use,
number tandem repeats. including 3 and 5 bases). Because unrelated people
almost certainly have different numbers of repeat units,
3. Satellite DNA: If the repeating unit is the length STRs can be used to discriminate between unrelated
between 100 to several thousands of base pairs then the individuals. These STRs loci (locations on a
sequence is called satellite DNA. chromosome) are targeted with sequence-specific
primers and amplified using PCR. The DNA fragments
DNA profiling process: The process begins with a that result are then separated and detected using
sample of an individual's DNA called a reference sample. electrophoresis. There are two common methods of
The most desirable method of collecting a reference separation and detection, capillary electrophoresis (CE)
sample is the use of a buccal swab, as this reduces the and gel electrophoresis.
possibility of contamination. When this is not available
(e.g. because a court order may be needed and not Each STR is polymorphic, but the number of alleles is
obtainable) other methods may need to be used to collect very small. The power of STRs analysis comes from
a sample of blood, saliva, semen, or other appropriate looking at multiple STRs loci simultaneously. The
fluid or tissue from personal items (e.g. a toothbrush, pattern of alleles can identify an individual quite
razor) or from stored samples (e.g. banked sperm or accurately. Thus STRs analysis provides an excellent
biopsy tissue). A reference sample is then analyzed to identification tool. The more STRs regions that are tested
create the individual's DNA profile using one of a in an individual the more discriminating the test
number of techniques, discussed below. The DNA becomes.
profile is then compared against another sample to
determine whether there is a genetic match. In practice, the risk of contaminated-matching is much
greater than matching a distant relative, such as
RFLP analysis: The first method for finding out contamination of a sample from nearby objects, or from
genetics used for DNA profiling involved RFLP left-over cells transferred from a prior test. For that
analysis (Restriction fragment length polymorphism). reason, multiple control-samples are typically tested in
DNA is collected from cells, such as a blood sample and order to ensure that they stayed clean, when prepared
cut into small pieces using a restriction enzyme. This during the same period as the actual test samples.
generates thousands of DNA fragments of differing sizes Unexpected matches (or variations) in several control-
as a consequence of variations between DNA sequences samples indicates a high probability of contamination for
of different individuals. The fragments are then separated the actual test samples. In a relationship test, the full
on the basis of size using gel electrophoresis. The DNA profiles should differ (except for twins), to prove
separated fragments are then transferred to a that a person was not actually matched as being related
nitrocellulose or nylon filter; this procedure is called a to their own DNA in another sample.
Southern blot. The DNA fragments within the blot are
permanently fixed to the filter and the DNA strands are PCR analysis: Developed by Kary Mullis in 1983, a
denatured. Radio-labeled probe molecules are then added process was reported by which specific portions of the
that are complementary to sequences in the genome that sample DNA can be amplified almost indefinitely. This
contain repeat sequences. These repeat sequences tend to has revolutionized the whole field of DNA study. The
vary in length among different individuals and are called process, the polymerase chain reaction (PCR), mimics
variable number tandem repeat sequences or VNTRs. the biological process of DNA replication, but confines it
The probe molecules hybridize to DNA fragments to specific DNA sequences of interest. With the
containing the repeat sequences and excess probe invention of the PCR technique, DNA profiling took
molecules are washed away. The blot is then exposed to huge strides forward in both discriminating power and
an X-ray film. Fragments of DNA that have bound the the ability to recover information from very small (or
probe appear as dark bands on the film. degraded) starting samples.

However, the Southern blot technique is laborious, and PCR greatly amplifies the amounts of a specific region of
requires large amounts of under-graded sample DNA. DNA. In the PCR process, the DNA sample is denatured

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 Singla et al. European Journal of Pharmaceutical and Medical Research

into the separate individual polynucleotide strands 6. Aggrawal A. Textbook of Forensic Medicine &
through heating. PCR uses replication enzymes that are Toxicology. 1st ed. New Delhi: Avichal publishing
tolerant of high temperatures, such as the thermo-stable company: 2014, p. 521-2.
Taq polymerase. In this fashion, two new copies of the
sequence of interest are generated. Repeated
denaturation, hybridization, and extension in this fashion
produce an exponentially growing number of copies of
the DNA of interest. Instruments that perform thermal
cycling are now readily available from commercial
sources. This process can produce a million-fold or
greater amplification of the desired region in 2 hours or
less.

However, the PCR method was readily adaptable for

analyzing VNTRs, in particular STRs loci. In recent
years, research in human DNA quantitation has focused
on new "real-time" quantitative PCR (qPCR) techniques.
Quantitative PCR methods enable automated, precise,
and high-throughput measurements. Inter-laboratory
studies have demonstrated the importance of human
DNA quantitation on achieving reliable interpretation of
STRs typing and obtaining consistent results across
laboratories.

DISCUSSION
DNA fingerprinting can conclusively prove the paternity
and solve cases explicitly. Thus, with the dawn of DNA
analysis and sequencing techniques, scientists
increasingly began to look at people's genomes when
questions of parenthood arose. DNA profiling is a tool
that is not only used to apprehend the guilty but also to
exonerate the innocent. As it often happens in the justice
delivering system, conventional evidence can be
tempered with, witnesses turn hostile, but DNA evidence
remains the same. The passage of time does not affect it
and neither does it change. DNA evidence thus unravels
the truth - it never lies. This approach proved
exceedingly useful; in fact, current marker-based
methods of analysis yield test results that are both
99.99% accurate and applicable in a variety of settings.
With the ongoing advancement of DNA sequencing and
analytical technologies, we will no doubt continue to see
an increase in the utility of these tests, as well as in the
availability of detailed genetic services to the general
public. Thus.

REFERENCES
1. Joseph W. The Blooding. New York: A Perigord
Press Book: 1989, p. 83.
2. Jeffreys A.J., Wilson V., Thein S.W. "Hypervariable
'minisatellite' regions in human DNA". Nature.
1984; 314: 67–73.
3. Joseph W. The Blooding. New York: A Perigord
Press Book: 1989, p. 202.
4. "Use of DNA in Identification" [Internet]. Available
from: Accessexcellence.org. Retrieved 2010-04-03.
5. Aggrawal A. Textbook of Forensic Medicine &
Toxicology. 1st ed. New Delhi: Avichal publishing
company: 2014, p. 525.

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