3.
DNA FINGERPRINTING (PROFIILING)
SYNOPSIS
1. Introduction **************
348
2. What is DNA? *****''***** 348
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3. Source of DNA * * * ' **** **** " *** ****
* ******* 349
.. 351
4. DNA Profiling Tcchnique ****** ******'******** ***
5. Prospects of DNA fingerprinting in India . . 352
******"*'***********. 353
6. Forensic Applications of DNA Test.
1. Introduction
In most of the crimes, establishing identity of an individual, be it
a victim of crime or the suspect, becomes the backbone of police
investigation. Fingerprint science has been, so far, the only infallible
means available to meet this requirement to a limited extent. The chances
of establishing identity on the basis of biological evidence, in particular,
genetically determined markers like blood groups and red cell enzymes,
have not yielded satisfactory results. Each one of the methods showed
relatively low exclusion power. Even the best efforts made by combining
all the known genetic marker systems, individualisation has always
remained elusive. But recently, DNA, which has a distinct genetic code,
has made a spectacular break through in biotechnology. To the forensic
scientist, it is the single most genetic marker and the most powerful tool.
now available, for individualising and discriminating the biological evidence
in forensic investigations. The term "DNA fingerprint'" was a trade mark
name given by Cellmark Diagnostics, a Company in America which
licensed the technique, developed in U.K. Its initial name helped to carry
the message conveyed by fingerprints to a lay man. The more appropriate
term now gaining currency is DNA Profiling.
2. What is DNA?
DNA is an abbreviated form of Deoxyribo Nucleic Acid. It is a basic
genetic material present in all nucleated cells of the body, and provides
the genetic blueprint of all life;
meaning thereby, it stores all hereditary
characters of an individual which he inherits from his
parents.
DNA is a linear double stranded chain of molecules which has a
helical structure, almost resemblng a twisted ladder with sides made of
348
DNA Fingerprinting (Profiling) 349
rope. Each DNA molecule consists of sugar (S) and phosphate (P) which
alternate together to support the rungs of the ladder. Whereas, four bases
namcly, adenine (A), thymine (1). guanine (G) and cytosine (C) join to
farn the rungs of the ladder. Adenine unites with thymine to form one
vpe of rung. and cytosine joins with guanine to form another. Because
of their specificity A will always pair with T andG will always pair
wth C.
The two strands are held together by hydrogen bonds between
complementary base pairs, Ifone strand showed AAGCTGA as a sequence
of base. the other would always be TTCGACT.
The structural units of DNA are called nucleotides. Each nucleotide
consists of sugar, phosphate and a nitrogenous base. The four different
DNA nucleotides will possess identical sugar and phosphate groups, but
they can only have either of the four bases. A molecule of DNA is
composed of four nucleotides linked to form a polynucleotide chain.
(Fig. 44)
Prof. Alec Jeffreys of Leicester University, England did pioneering
work in developing DNA fingerprinting technique which is now a days
widely accepted technique for forensic investigations. He observed that
while large sections
of DNA show similar base sequence for every human
being, they
as are made of same body
components, a few sections
human DNA found varying from individual to individual.
are
of
This
observable variation in human DNA is called
polymorphic segments in the DNA molecule serve polymorphism.
These
individuals. On the basis of these
as a means to
identify
for mono
findings, it was revealed that, except
zygotic twins, each individual can be differentiated from another.
3. Source of DNA
DNA in each individual is the same
in each and every human cell,
but it differs in every individual. For DNA
1s the DNA profiling, the basie material
itself. To determine the
extracted from the
individuality, the DNA has to be first
specimen. It can be extracted from any tissue or body
uids. It is found
present in every living cell of our body, such as blood,
Semen, hair roots, bone (marrow), fresh
tissue, an autopsy specimnen.
sallva, urine etc. Finding of polymorphic chain is, however, easy when
ne iresh specimen is drawn. Saliva and urine
may contain negtngtoic
guantity of DNA., and therefore, DNA profiling may pose probiems
Forensic Science in Crime
Investigation
//S/e/7e4
A=T
GC
T=A-
G C
TP
Fig. 44 DNA double nelix structure with alternating
Phosphate & Sugar units and base pairs of (Adenine
Thymine, A-T: Guanine Cytosine, G-C)
DNA Fingerprinting (Profiling) 351
DNA
ProfilingTechnique
4.
e DNA profiling technique involves a lengthy
involved:
procedure. Following
the steps
are
1 Extraction and purification of DNA from
specimen.
Fragmentation of DNA using "restriction enzyme".
3Arranging the fragments on the basis of their length by agarose
gel electrophoresis.
(4) Transfer of separated fragments to nylon membrane by "Southern
blotting
(5) Hybridisation with a radioactive labelled probe.
(6) Visualising the bands of DNA by
autoradiography.
A brief procedure for DNA
profiling is given below:
DNA is first extracted from a
specimen by an elaborate chemical
process. It is then mixed with a special
enzyme called "restriction
endonucleasis"'which function as molecular scissors a sort of
biological
-
tool to cut the DNA at specific sites. The
number and length of the DNA
fragments depends on where and how often the
occurs in the DNA
enzyme's base sequence
specimen. This fragmentation in length as well as in
number varies from DNA
specimen of one individual to another.
The
technique of analysis is known as restriction
polyphormism (RFLP). The restricted fragments are fragment length
then separated
according to size using gel electrophoresis
put on a gel and an electric technique. The fragments are
current is applied. The shorter
gel faster towards the positive pole than the fragments
across the move
This separates the DNA longer fragments.
fragments on the basis of their length.
The next
step is to transfer the DNA
nylon membrane the fragments from the gel to a
,
process called as ""Southern blotting". This process
Ixes the DNA
his is
fragments to the membrane firmly and in the same position.
followed by hybridisation
COntact a radioactive labelled process. Hybridisation involves bringing
nolecule that binds
probe, which is a particular type of DNA
specifically with its complementary base sequence
present in the
membrane. The probe bound fragments, being radioactive,
a n then
be recorded on an
X-ray plate, the process known as
auLOradiography. The X-ray plate will show dark bands appearing very
352
For
orensic Science
in Crin
rime Investigatior
much like bar
The code. This, in profiling
comparison of h utshell, 1sis the
utshell, tne process of DNA
from the same ands the bands in the DNA
DNA profilesaf differen
of different specimnen
of
pro
individual will show similarity, whereas,
as. DNADNA spspecimen
Cnt i t
individuals will show different DNA band protiies
More
recently,
PCR) has been another technique known as polymerase is specially
chain reaction
developed for DNA profiling. This method specially
recommended where DNA is insufficient for analysis or itDNA
is in SEgments
degraded
O. The PCR technique takes into account
PcSent in
polymorphic DNA Segmens
the
specimen, and by polvmerase chain reaction (PCR) produce
d p l e copies of a particular polymorphic DNA segment, suficent to
meet ne
t requirements of DNA profiling test. This method is particulariy
usetul to profile a single strand of hair, by extracting DNA from hair
TOcle cells, attached to the root. The PCR method, which takes into
account the sequence variation in small sections of DNA, can be used
tO Duild a computerised database for more objective interpretation or the
results.
5. Prospects of DNA ingerprinting in India
DNA profiling technology has been' widely used in the west
particularly in Britain, USA and Australia, with remarkable success. The
technique promises positive identification or elimination of suspect, on
the basis of specimens collected at the scene of crime. DNA profiling has
already caught the curiosity of forensic institutions in the country, the
police as well as the judiciary. The first case, which hit the headlines in
the Indian news papers, is the one presented to the Madras High Court
by the Experts' from the Tamil Nadu Forensic Science Laboratory. They
examined the blood samples in a paternity dispute involving a five year
old Mary in 1989.
In addition to HLA typing which is routinely done in the laboratory
their findings were conlimed by DNA profiling test caried at CCMB.
Hyderabad. The evidence was admitted by the High Court Judge, a first
case of its kind in the country. Only recently the Supreme Court in Indta
also suggested DNA test to prove the natural parenthood of the child,
claimed by two women wilh a common husband. It is evident that DNA
profiling has potential legal applicalion and of enormous utility in crime
invistigations
DNA Fingerprinting (Profiling) 353
The Department of Biotechnology, Home Ministry and the Council
Scientific and Industrial Research (CSIR) joined together to set up an
anomous DNA profiling centre, most likely in Hyderabad. The Centre
auto
far Ccllular and Molecular Biology (CCMB) whose scientists have
deycloped an indigenous DNA probe, and tested already in as many cases,
vould become the nodel agency for the centre. Some of the objectives
of the proposed centre are:-
(1) To provide
DNA fingerprinting facilities to crime
investigating
agencies in the rape and murder cases, including identification
of mutilated and unidentified bodies.
(2) To prepare and maintain DNA
profiles of rapists, criminals and
terrorists, to assist police personnel, forensic scientists,
and the judiciary for use in crime and
lawyers
family litigations.
(3) To interact with INTERPOL, keep DNA fingerprint records of
international criminals, provide centre's facilities to INTERPOL
or other nations, to facilitate
crime investigation and training.
(4) To cary out DNA profiles and related analysis in civil cases like
paternity disputes, immigration and exchange of new borns in
hospitals, to various agencies including private parties on payment
basis.
6. Forensic
Applications of DNA Test
DNA fingerprinting
technology has the potential to revolutionise the
identification process. It will help solve
many serious crimes, such as
sexual assault, homicides, paternity disputes etc., as
discussed below
Blood and semen in rape cases.
sexual offences, which traditional
Difficult problems connected with
be solved by DNA
serological tests could not solve, can
profiling test The blood stains, seminal stains and
.
vaginal swabs can be subjected to DNA profiling for
identifying the
or
eliminating the suspect with near certainty. Specimens collected rapist
from
one or more
suspects in the crimes like rape and murders or gang
can be scrutinised
,
rape
through DNA test to determine involvement or one,
more or none of the
suspects, and to provide investigative leads to the
police to nab the right suspect. In a
a
speciman of vaginal swab, which is
mixture of vaginal fluid and seminal
wIth ease, and the semen can be
component, the two ean be seperated
subjected to DNA specitic test.
FS-23
354 Forensic Science in Crime Investigation
Other bodymaterial: In assaults, hit-and-run accidents or homocidal
found at the scene ( such as
crimes, any type of body sample may be
the DNA test can identif
hairs, body tissues, saliva, teeth, bones, etc.)
be insufficient in hair or saliva
the person positively. Though DNA may
useful evidence
specimen, application of PCR technology may provide
extracted from the
skeletal
Unidentified dead bodies DNA has been
murder and identified with
remains of the 8 year old child, a victim of
bone DNA thus established the
the presumptive parents. The analysis of
forensic investigations.
feasibility of DNA typing of bones in
of the deceased can be
Likewise, postmortem tissues or organs
of the deceased
identified by DNA test to establish the likely parentage
in
is an issue many civil
Paternity disputes: Paternity determination
involving wide range of disputes like maintenance suits,
litigations out ofproperty
establishing parenthood disputes, inheritance disputes arising
inheritance or immigration problems. DNA test
is capable of resolving
most of these issues. DNA fingerprinting
has been regularly employed
on the basis of their
in Britain to screen immigrants claiming citizenship,
relations holding British citizenship.
Old Cases : Old and closed cases can be reopened in the light of
solution
DNA profiling capabilities, to re-evaluate the evidence that eluded
by conventional methods.
