AMITY INSTITUTE OF FORENSIC SCIENCE,

AMITY UNIVERSITY OF UTTAR PRADESH, NOIDA

NTCC TOPIC: DNA FINGERPRINTING TECHNIQUES

Submitted in partial attainment of the requirements for the grant of degree of

B.Sc. in

Forensic Science

Batch: 2018-2021

By:

PREETI THAKUR

A5905918230

Supervisor:

Dr. AMRITA DAS

Assistant Professor Amity Institute of Forensic Science

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 DECLARATION

I Preeti Thakur solemnly declare that the project report entitled “DNA Fingerprinting
Techniques” which is submitted by me to the AIFS, is partial attainment of requirement for the
grant of the degree of BSc. (H) in Forensic Science, has not previously devised for the basis for
the grant of any degree, or other alike kind of titles.

Theauthor attests that permission has been granted for the use of any copyrighted
material performing within the Dissertation/Project record other than brief excerpts requiring
best proper acknowledgement in scholarly writing and all such use is recounted.

Preeti Thakur

A5905918230

(2018-2021)

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 CERTIFICATION

On the basis of declaration by Preeti Thakur Enrollment number– A5905918230,

student of Forensic Science has completed work presented in the project of the term Paper
entitled “DNA Fingerprinting Techniques.” as a part of the Second Year program of BSc. (H)
in Forensic Science from Amity University, under my direction.

Dr Amrita Kumari Das

(Faculty Guide)

Department of forensic Science

AIFS, Noida, (UP)

_________________

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 CONTENTS

S.N CONTENTS PAGE NUMBER

o
1. Abstract. 5

2. Introduction. 6

3. Review of literature. 9

4. Methodology 10

5. Observation and Result 15

6. Conclusion 18

7. Scope of study 19

8. References 20

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 ABSTRACT

One of the major discoveries of the 20th century, DNA fingerprinting, has revolutionized forensic
investigations. This review briefly summarizes 30 years of progress in the study of forensic DNA
that helps to convict suspects, exonerate the falsely convicted, and locate victims of crime,
catastrophes, and war. Present standard approaches are covered, based on short tandem repeats
(STRs) as well as lineage markers (Y chromosome, mitochondrial DNA), and casework examples
demonstrate applications. There is a discussion of the merits and risks of expanding forensic DNA
databases and we wonder what the future contains for forensic DNA fingerprinting.

The primary aim of forensic DNA fingerprinting is to invent absolute certainty beyond a reasonable
doubt that the questioned samples and reference samples are linked. This complex and durable
technique, however, necessitates a set of methodologies and protocols that forensic scientists must
follow in order to achieve an accurate result. DNA fingerprinting techniques have accelerated as a
result of a series of significant technical advancements. It started with a technique called "restriction
fragment length polymorphism (RFLP)," which progressed through the phases of "minisatellites," or
VNTR analysis, and is now moving through the stages of "microsatellites," or STRs and SNPs.
Similarly, the instrumentation processes moved from horizontal to vertical gel systems to automated
analyzers with responsive microcapillary arrays and laser-based detection systems. For data analysis
and the development of user-friendly reports, high-quality software is now available.

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 INTRODUCTION

DNA FINGERPRINTING IN FORENSICS: PAST, PRESENT & FUTURE.

The past - a new method that changed the forensic world

Alec Jeffreys discovered at Leicester University, UK, that repetitive DNA patterns studied with
multi-locus probes are remarkably variable and genetically inherited. He refrained from naming the
process 'DNA fingerprinting' after himself. His discovery opened a new branch of science under this
name. The technique has proven to be applicable in many biological disciplines, namely in research
on species diversity and conservating, as well as in clinical as well as anthropological studies.

Forensic genetic fingerprinting can be described as a comparison of the DNA in nucleated cells of an
individual with that discovered at the scene of a crime identified in biological matter or with the
DNA of another person for identification or exclusion purposes. A young boy was rescued from
deportation by the first application of DNA fingerprinting and the process thus won the sympathy of
the public. The time in the 1990s was the golden research era of DNA Fingerprinting, he writes,
succeeded by two decades of engineering, implementation, and application of high-throughput. A
mood of collaboration and interdisciplinary research, he says, characterized the years after the
uncovering of DNA fingerprints. “An interdisciplinary mood of study and collaboration
characterized the years following the discovery," he adds. The author is a professor at the School of
Oriental and African Studies at the University of Cambridge. 

The initial technology of Jeffreys, now obsolete for forensic use, underwent major advances.
Spectacular fallacies exposed significant insufficiencies in the technological protocols, from the
1989 case of People vs. Castro in New York to the case against Knox and Sollecito in Italy. DNA
proof is an important aspect of factual evidence nowadays and requires close examination. Of
course, genetic fingerprinting in some of the many countries in the world that use this tool will not
decrease the crime rate. DNA profiling, however, adds harsh scientific value to the facts and thereby
enhances the justice system's legitimacy.

The technological evolution of forensic DNA profiling

Radio labeled DNA probes containing minisatellite or oligonucleotide sequences are hybridized into
DNA that has been digested with a restriction enzyme in the classical DNA fingerprinting process.
The most frequently used minisatellite probes, numbered 33.6 and 33.15, were in the UK, most parts
of Europe and the USA. These so-called multilocus probes (MLP) detect sets per person of 15 to 20
variable fragments ranging in size from 3.5 to 20kb. But despite its popular application to crime and
kinship cases until the middle of the 1990s, the multi-locus profiling approach had some constraints.
A DNA code, which could ideally be produced even from a single nucleated cell and highly
degraded DNA, was required. This code, say the researchers, could be quickly produced,

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numerically encrypted, compared automatically, and easily supported in court. Only 10ng of
genomic DNA is required for the method and has been validated through comprehensive
experiments and forensic casework.

STR typing is more sensitive than the RFLP methods of single-locus. Currently, forensic DNA
profiling is achieved using a multi-allelic STR marker panel. The chance that two individuals would
have similar markers within their DNA at each of 13 separate STR loci exceeds one out of a billion.
Up to 30 STRs can be found in a single injection of capillary electrophoresis, producing a unique
genetic code for each person. The application of these markers has been enhanced by the
introduction of these STR markers into commercial kits. Government-controlled DNA databases for
criminal investigation were first developed in the UK in 1995. With approximately 6 million
accounts, the UK maintains the biggest DNA database in the world. Approximately 10% of the
population in the UK is in the national DNA database, while the figure in Germany and the
Netherlands is only about 0.9% and 0.8% respectively.

Lineage markers in forensic analysis

In cases where there is excess DNA from a female victim and only a low proportion from a male
suspect, Y chromosome analysis is useful. For the study of low-level nuclear DNA samples,
mitochondrial DNA (mtDNA) is of interest. The discovery of two missing children of the Romanov
dynasty, the last Russian monarchy, is a classic example. Big, representative, and quality-assessed
databases of haplotypes sampled in appropriate reference populations are needed to determine the
match probability between Y-STR or mtDNA profiles using the most commonly used counting
process. Other studies have shown that it is possible to trace very distant family branches back to
common ancestors who were living hundreds of years ago.

The existence in the mtDNA of the Tsar and his family, which was a controversial finding in 1991,
of two slightly different mtDNA haplotypes within an individual, is now understood to be relatively
common and can be checked at certain positions for its frequency. YHRD (www.yhrd.org), hosted
by the Institute of Legal Medicine and Forensic Sciences in Berlin, Germany, is the largest forensic
Y chromosome haplotype database. In Innsbruck, Austria, the largest forensic mtDNA database is
EMPOP, with about 33,000 haplotypes in 63 countries. In non-suspect cases, this link between
genetics, genealogy, and geography could provide investigative leads for investigators. Knowledge
on the bio geographical roots of unknown DNA could also be collected from a variety of informative
SNPs of ancestry.

Benefits and risks of forensic DNA databases

The steady increase in the size of the database of forensic DNA raises concerns about the inclusion
and retention requirements. Family DNA Database Searching is based on close matches between a
crime stain and a data-based person, which may be a true perpetrator's close relative. The first
successful family search was undertaken in the UK in 2004 which resulted in Craig Harman being
convicted of manslaughter. For democratic societies, human rights and freedoms are critical and

7
attempts to expand forensic DNA databases to whole populations need to be condemned, writes
Peter Bergen. After a big setback at the European Court of Human Rights in 2008, he writes, the
improvements to the UK database came in the 2012 Security of Freedoms bill. A DNA database
containing samples from every Portuguese citizen was proposed by the incoming government of
Portugal in 2005, but it was limited to offenders. It is important to follow common ethical and
privacy principles for DNA database creation and governance.

The future of forensic DNA analysis

An increasing number of colleagues are persuaded that approaches focused on fragment length
analysis would soon be replaced by DNA sequencing. Next Generation Sequencing (NGS)
technologies can theoretically be rapidly and cost-efficiently extended and analyzed. The prospect of
on-site DNA analysis in the US has already changed the way DNA can be gathered in the future.
Driven by rapid technological development, DNA is simply another indicator of rapid identification,
writes Mark O'Mara. In Germany, for example, for identification purposes only, the use of DNA
profiling is prohibited in criminal proceedings, he writes. O'mara: It remains to be seen if quick
DNA technologies can change the way police collect DNA in other nations, he adds. The lives of
millions of people worldwide are directly affected by forensic DNA technology. At present, a
worldwide framework for interviewing DNA profiles from criminal justice databases continues to be
a very distant project. CSI watchers know, and even experts assume, that the case will eventually be
solved by DNA only after the slogan Don't Ask, it's DNA, dumb! It should not be concluded that the
social and ethical costs would inevitably override the advantages of forensic DNA fingerprinting.

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 REVIEW OF LITERATURE

UNDERSTANDING THE BASICS OF DNA FINGERPRINTING

In the field of human genetics, scientific technology has recently made important advances. The
discovery of deoxyribonucleic acid (DNA) was a watershed moment in scientific history. Dr.
Alec Jeffreys discovered that some DNA regions contained DNA sequences that were isolated
repeatedly next to one another. He also discovered that the amount of repetitive parts in a sample
can vary from person to person, which is why it's useful in forensic science. Though the
application of such information is not yet completely understood, the subject has unlimited
potential. The purpose of this paper is to set out the groundwork for understanding the various
methods of DNA fingerprinting and their applications in forensic science.

DISCOVERY, DEVELOPMENT AND CURRENT APPLICATIONS OF

DNA IDENTITY TESTING
In genetic identity verification, DNA fingerprinting, DNA profiling, and DNA typing are all
used. About 99% of the DNA sequences in the human genome are similar from one individual to
the next. To differentiate all humans, a limited number of sequence variations are used.

Sir Alec Jeffreys established DNA-based identity testing in the United Kingdom 20 years ago.
His team created a short-sequence radioactive probe that could lock onto repeating sequences.
According to Jeffreys, repeating sequences "may be highly variable, insightful genetic markers."
Each individual has a signature fingerprint, which is similar to a human family's DNA
fingerprint. Each one had a different pattern of DNA segments, which were made up of 15 to 20
bands. However, a closer examination of the human family's patterns showed that the mother and
father both had their own pattern, with the infant having a combination of both. Professor
Jeffreys and his colleagues published their first paper on DNA fingerprinting in the spring of
1985. The results were published in newspapers.

DNA FINGERPRINTING: HELPING HAND IN SOLVING CRIME

The advent of DNA fingerprinting aids in the investigation of crimes, and our Indian system is
not oblivious to this. The Criminal Code of Procedure, 1973, was revised in 2005 to authorise
medical professionals to examine anyone detained on suspicion of committing a crime. In civil
cases, DNA analysis is also critical in establishing the paternity of a child. In India, the use of
this evidence is particularly important in criminal, civil, and identity theft cases.

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 METHODLOGY
There are some basic steps which needed to be followed for DNA fingerprinting are as follows:-

I – DNA ISOLATION

• The isolation of DNA should be done from cells or from tissue.

• For this purpose of isolation, only a very small amount of sample like blood ,hair etc is
required.

II – DNA DIGESTION

• The digestion of DNA can be performed with the help of restriction endonucleases.

• Then, the DNA with the help of restriction enzyme is cut into fragments.

• Certain restriction enzymes are used for DNA cutting and each restriction enzyme cuts the
DNA at specific region or sequence.

• This specified cutting sequence of DNA are said to be “ restriction fragments”.

• There are some immensely used restriction enzymes used are HaeIII , Alu I etc.

III – SEPARATION OF DIFFERENT DNA FRAGMENTS

• The separation of DNA fragments, on the basis of their size is done by with the help of the
process called “Gel Electrophoresis.”

IV – TRANSFERRING OF DNA FRAGMENTS

• The fragments or the pieces of DNA are shifted to nylon sheet or nitrocellulose membrane.

• This nylon sheet is then placed on gel soaked them overnight by the process of “ Southern
Blotting”.

V – PROBING AND LABELLING

• The addition of radioactive probes to nitrocellulose membrane, which are complimentary to the
targeted sequence.

• This probe only sticks to specific region on the nylon membrane.

VI – HYBRIDISATION

• The DNA which is labelled, must be hybridised with complimentary sequences, present on
nitrocellulose membrane. So that their position can be identified for later .

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• Then this particular membrane is washed off for the removal of extra probes , also making the
background clear.

VII – AUTORADIOGRAPHY

• To identify the places where radioactive probes binds to the fragments of DNA.

• These identified marks are known as “Dark Bands”.

• After all this , an X- ray film is exposed to nitrocellulose membrane.

• And this process is said to be autoradiography.

VIII – ANALYSIS OF BAND PATTERN’S

• The band’s position of different individual’s are compared.

• There are various computer software present for this purpose.

TYPES OF DNA FINGERPRINTING METHODS

The DNA Fingerprinting can be done with the help of these techniques are –

a) Electrophoresis.

b) Polymerase Chain Reaction (PCR).

c) Restriction Fragment Length Polymorphism (RFLP).

d) Random Amplified Polymorphic DNA (RAPD).

e) Amplified Fragment Length Polymorphism (AFLP)

a) ELECTROPHORESIS
• Arne Tiselius is considered as the father of this technique.

• It is a technique of separation that depends upon the movement of ions under the influence of
applied electric field.

• The major principle behind this technique is that the ions which carries positive charge will
move towards negative electrode whereas the ions carries positive charge will go at negative
electrode.

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• Now, the mobility of an ion is depend on various factors such as-

a) Size and Shape Of The Molecule – Helps in assessing the particle’s velocity under which they
starts moving in applied electric field.

b) Strength of Electric Field - The strength of electric-field can be determined by this formula –
F = QV. The force applied on tbe particle and charge will helps in the determination of electric
field. When the force is applied, a particle will starts moving but the movement us also opposed
due to the frictional force acting because of velocity.

b) POLYMERASE CHAIN REACTION

• This technique was developed by Kary Mullis in the year 1983.

• It is a process or method which is used to make multiple copies of DNA segments with tiny
amount of sample of DNA and then analyse it to make several copies.

• There are three main components required for carrying out this technique are as follows-

a) DNA Template – This is also known as DNA Target Sequence. It is that molecule of
DNA which have that region of DNA which is to be amplified.
B) DNA Polymerase – This Polymerase works by adding nucleotides, complimentary to the
template strand and the new stranded DNA is complimentary to this targeted sequence. The
commonly used Polymerase is Taq Polymerase.

b) Primers – These are the sequence of a short nucleic acids. They provides a starting point
for the synthesis of DNA. The primers are of two types – Forward and Reverse Primers.

STEPS OF PCR-

1. DENATURATION-
• The heating of reaction mixture is carried out for one minute at a temperature of 94°c.

• This helps in the separation of double stranded DNA.

• The separated DNA will acts as a template for the synthesis of DNA.

2. ANNEALING-
• After Denaturation, the cooling of reaction mixture is done at a temperature of 54°c .

• This results in the separation of primer to separate the strands of the DNA.

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3. EXTENSION-
• The reaction mixture is again heated at a temperature of 72°C which is ideal temperature for the
working for Taq Polymerase.

• This adds the nucleotides, which are complimentary to primers.

• Thereby there is a extension of new strand.

NOTE- These three steps of PCR are repeated for 30- 40 cycles for the production of million of
exact copies of targeted DNA.

c) RESTRICTION FRAGMENT LENGTH POLYMORPHISM (RFLP)

• This helps in the length analysis of the molecule of stranded DNA by using repeating base pair
patterns.

• This process is carried out by taking the sample of fragmented DNA and using restriction
enzyme, this enzyme can recognize and cut the DNA wherever a specific short sequence
happens, called as restriction digest.

• Then, the resulted fragments of DNA are differentiated by length with the help of agarose gel
electrophoresis and later shifted to a membrane by Southern Blotting.

• Then this membrane is hybridised to labelled DNA probe which identifies the fragment length,
and are complimentary to probe.

• An RFLP takes place if the length of detected fragment differs among individuals. Every
length of fragments is an allele, which can be used for analysis of gene.

d) RANDOM AMPLIFIED POLYMORPHIC DNA (RAPD)

• This is a type of PCR reaction, but the DNA segments are amplified at random.

• It makes various short primers and then PCR is performed by taking a large template of
genomic DNA, so that the amplification of fragments can occur.

• After resulting patterns, a semi-unique profile is obtained from this RAPD reaction.

• RAPD does not depends upon the knowledge of the DNA sequence of the target organism: the
identical 10-mer primers may or may not amplify the DNA sequence , depends on the positions,
complementary to the sequence of primers.

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e) AMPLIFIED FRAGMENT LENGTH POLYMORPHISM
• It is a faster technique than analysis which uses PCR for amplification of the samples of DNA.

• This depends on variable number tandem repeat (VNTR) polymorphisms for the differentiation
of various alleles, which were separated on a poly-acrylamide gel.

• With the help of PCR analysis for amplification of the mini-satellite loci of human cell, this
method was faster in recovery than the RFLP.

• However, use of the gel in the analysis phase, there is trouble in bunching of the VTRN’s,
which causes misidentifications during this process.

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 OBSERVATION AND RESULT
DNA fingerprinting has established to be a powerful and effective tool in solving crime and
putting the criminal responsible for a particular crime behind the bars. DNA fingerprinting is
being used in forensics since many years and has improved the field 10 folds.

From immigration cases to solving gruesome serial killings DNA-fingerprinting has been used in
various cases some of which will be discussed following:

The first case: an immigration dispute

A Ghanaian family immigrated to the UK and became citizens. However, one of the son returned
to Ghana and was denied entry to the United Kingdom due to a counterfeit passport. Professor
Jeffreys was approached by the family's lawyer, who asked if he could confirm that the boy was
mother's son and not her nephew (she had several sisters in Ghana). The absence of the father
complicated the situation. The mother, the son whose identity was contested, and the mother's
three undisputed children all had DNA samples taken. The patterns reinforced the mother's
relationship with the son in question. Furthermore, the tests revealed that all four children were
born to the same parent. This immigration case asphalted the way for DNA fingerprinting to be
used in forensic cases and for determining identity.

Refining the assay

Professor Jeffreys was flooded with questions following the case's progress. In 1986, he got a
phone call from police in Leicestershire, England, asking for his assistance in solving a double-
murder case. Because of the large amount of DNA material needed for the test to be accurate,
Professor Jeffreys claimed his initial DNA fingerprinting methods would not work in a criminal
case. It will be difficult to know where the DNA bands are approaching from and which
minisatellite regions are involved if the amount of DNA evidence is limited and only 15 to 20
DNA bands are examined. The procedure is very time-consuming. Professor Jeffreys began by
using a probe with sequences that bind to different minisatellite loci. He created a probe that
latches to a single minisatellite locus to simplify his DNA fingerprinting assay. A single-locus
probe can only detect two DNA segments in a person, each of which corresponds to two alleles:
one inherited from mother and one from father. In 1986, Professor Jeffreys used this new method
to solve a double-murder case.

Identifying a killer

Two 15-year-old girls were raped and murdered in Leicestershire. In spite of the fact that the
attacks happened three years apart, the similarities led the police to conclude that the perpetrator
was the same person. The case has become even more perplexing after a suspect in custody
admitted to the most recent murder but not the previous one. Professor Jeffreys was asked to
perform DNA profiling on a blood sample taken from the perpetrator, as well as tissue samples

15
and sperm from the two victims. The DNA profiling showed that both victims' sperm were
similar, suggesting that both murders were committed by the same male. In an unexpected twist,
the findings also revealed that the suspect in police custody was not the killer, allowing him to be
released and being the first suspect to be convicted of a crime using DNA evidence.

The killer's DNA profile was then matched by a large-scale manhunt to find the individual whose
DNA profile matched that of the killer's sperm. Both adult men in the neighborhood were asked
to provide blood or saliva samples for examination. More than 5000 samples were collected, and
DNA profiling was performed on the 10% of men who shared the killer's blood type, but no
match was made. The fact that this modern and advanced test failed to locate the killer
disappointed both the police and the public.

A woman confirmed overhearing a man claim to have provided blood on behalf of a colleague,
Colin Pitchfork, six months after the initial inquiry. Pitchfork was apprehended and his blood
checked, yielding the long-awaited DNA match, and Pitchfork was found guilty of both murders.
As a result of this promising outcome, DNA profiling has become a powerful tool in the
investigation of crimes.

Newer methods and the “Angel of Death”

DNA profiling was being used all over the world by the end of 1986. With the invention of the
polymerase chain reaction (PCR) and the discovery of various repeating sequences known as
microsatellites, DNA typing was refined. DNA amplification by PCR improves sensitivity,
allowing even archival and partially degraded samples to be analysed with small amounts of
DNA.

Mini-satellites, also known as variable numbers of tandem repeats (VNTR), are repeating
sequences of unit lengths ranging from 6 to 100 bases. At each mini-satellite, these units can be
replicated two to several hundred times. Thousands of different minisatellites are strewn about
the genome, but they tend to congregate near the telomere, or chromosome's end.

Microsatellites, also known as short tandem repeats (STR), are made up of a unit that can be
anywhere from one to seven bases long. At each microsatellite locus, this unit is replicated 5 to
100 times. Thousands of different microsatellites are dispersed across the genome at random,
rather than in a single area.

The 40-year search for Nazi prison camp doctor Josef Mengele, who fled from the Allies at the
end of World War II, was brought to an end by PCR-based DNA typing. Mengele, dubbed the
"Angel of Death" at Auschwitz, was believed to have fled to South America. Mengele drowned
at sea in 1979 and was buried in Brazil, according to a tip to the police. In 1985, the badly
decomposed remains were exhumed in order to obtain DNA samples, but the specimens were so
weak that Professor Jeffreys resorted to reverse paternity testing, reconstructing Mengele's DNA

16
pattern using blood samples from Mengele's wife and son. The remains of Mengele were
assumed to be his in 1992.

Development of a DNA database

The DNA Identification Act of 1994 allowed the FBI to extend a pilot project into a national
DNA database, the Combined DNA Index System, as a tool for investigating violent crimes in
the United States. CODIS incorporates DNA analysis with computer technology to enable local,
state, and national crime laboratories to electronically share and compare DNA profiles.

The Forensic Index, which contains DNA profiles from crime scene evidence, and the Offender
Index, which contains profiles from those convicted of felony sexual crimes and other violent
crimes, are the two indexes in the database. The mechanism is focused on the amplification of 13
core STR loci as well as the amelogenin gene, which is found on both the X and Y chromosomes
and can be used to determine a person's sex.

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 CONCLUSION

DNA fingerprinting is a most reliable way to identify living organisms. DNA is a distinguishing type
of genetic material in organisms, having caracteristics of a kind. DNA is not simple to be altered
after it is left at the location of a crime scene or present with a mummy, this makes it a valuable
forensic tool. RFLPs and VNTRs are the oldest techniques of DNA fingerprinting, which uses a
large numbers of sample which uses the probe hybridization methods for the detection of
polymorphism in the DNA. STRs are the present form of DNA fingerprinting, and is based on PCR,
uses a very small sample of DNA. DNA fingerprinting has many uses including criminal rape cases,
paternity tests, molecular archeology, sports memorabilia, etc. In DNA ,there are no two exactly
alike molecules, but is one of the only things in common in biological organisms are made with.
DNA forensics is the most useful tools in linking together a crime scene. In past few years, there are
many advanced methods for the collecting and preserving of DNA samples to help the reliance of
this evidence in the court room. Advanced techniques and storing facilities to prevent the
degradation of evidences, forensic science has achieve a huge progress in allowing DNA samples as
strong evidence in courtrooms. DNA evidence is presently one of the most strong tool used to solve
the crime scene. The possibilities of where criminals altering their fingerprints and other physical
evidences, DNA evidence is one of the only strong methods used for the identification purposes. The
chemicals namely luminol, can be used for crime scene analysis in such cases where initially with
the help of naked eyes it is not possible to identify or detect the presence of any kind of evidences
like blood drops etc but when luminol is sprayed over the surface , we can easily identify or see the
evidence . Although, there are various factors that makes it hard for preserving a good DNA sample,
progress have to be made forensic science field, and further seems to have a limitless and broad
future in technology to come.

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 SCOPE OF THE STUDY
 Over the last few years, there has been many changes in DNA collection methods which
made it possible to present DNA as evidence in court in front of jury.
 Standardisation have been done related to various aspects of crime scene investigations,
which create persistent with all kind of evidence.
 The only basis DNA is now accepted in the court is since of techniques which enables the
proper DNA collection, transportation, analyzation, and storing of DNA sample without any
degradation.
 When DNA sample is placed in its natural state it becomes the most relevant evidence in
court.
 When a crime scene is examined, sometimes it seems that the culprit got away without
leaving any evidence behind. In previous years, violent crimes have taken place and cleared,
leaving no visible evidence related to the crime or crime scene.
 It is specificaly clear or we know that small biological fluids particles like blood can adhere
to various surfaces of the crime scene for a long period of time without anyone ever noticed
but they are there for that time.
 But there is with the changes along time a chemical that helps us to visible the microscopic
blood particles and find the proofs with naked eyes it is impossible to see these evidences.
 The name of this chemical is luminol which helps in revealing the particles with a light-
producing chemical reaction between hemoglobin and several chemicals.
 The reason behind the production of this light because the reactants have more energy
reactants and liberate extra energy in the form of light photons. This process is also
responsible for the glow of fireflies and known as chemi-luminescence.
 After the chemical is sprayed on the questioned sample, a blue glow will appear in the part
where there is the presence of blood which helps in generating the strong evidence and helps
in solving of the heinous crimes.
 In future, new techniques of visualization will be devised that helps in continuing the
structure of DNA.

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