DNA Profiling

Aims & Objectives:

DNA profiling (also called DNA Fingerprinting or DNA Typing) is the process of determining
an individual's nucleotide sequences of certain area of DNA, which are as unique as fingerprints.
Each person has a unique DNA Fingerprint. Unlike a conventional fingerprint that occurs only on
the fingertips and can be altered by surgery, DNA Fingerprint is the same for every cell, tissue
and organ of a person. It cannot be changed by any known treatment. The Ideal way to
distinguish an individual from other people would be his or her entire genomic DNA sequence.

Review Of Literature:
The study of finger, palm and sole prints is called Dermatoglyphics. It has been a subject of
human interest since primitive times when man used to hunt for his food with the help of animal’s
foot prints. Science of fingerprinting was first used by Sir William Herschel as a method of
identification in 1858. In India the science of fingerprints was discovered by chance during a
murder investigation in Jalpaiguri in 1897.
Alec Jeffreys, a British genetist, invented the DNA Fingerprinting Technique at Leicester
University, United Kingdom. He used VNTRs as genetic markers. He developed a scheme to
measure the length of DNA fragments containing the repetitive sequence. It is also known as
Restriction Fragment Length Polymorphism (RFLP).
Dr V.K. Kashyap & Dr Lalji Singh started the DNA Fingerprinting technology in India at CCMB
(Centre For Cell And Molecular Biology) Hyderabad. In this technique radioactive DNA probe is
obtained from Y Chromosome of female banded krait snake (in this snake female has XY and
male has YY chromosomes). The unique segment obtained from this chromosome is Banded
Krait Minisatellite (BKM-DNA).

Material & Method Used:

Techniques For DNA Fingerprinting-
1. DNA Isolation - The DNA is extracted from the nuclei of white blood cells of
spermatozoa or of the hair follicle cells that cling to the roots of hairs that have fallen, or
been pulled out.
2. DNA Fragmentation - The DNA molecules are first broken with the help of enzyme
restriction endonuclease (called chemical knife or molecular scissors) that cuts them
into fragments. The fragments of DNA also contain VNTRs.
3. DNA Amplification - Fragments of a particular size having VNTRs are multiplied through
PCR technique. They are treated with alkaline chemicals to split them into single
stranded DNAs.
4. Electrophoresis - The DNA samples are loaded on Agarose Gel Electrophoresis. The
charge is applied. The DNA fragments, which are negatively charged move to the
 positive pole. The movement of these fragments depends on the length of these
fragments. This results in formation of bands.
5. Southern Blotting - The separated fragments of single stranded DNA are blotted onto a
Nylon Membrane (Nitrocellulose Paper). The single stranded DNA gets embedded into
nylon membrane. This technique is named after the inventor, Edwin Mellor Southern
(1975).
6. Hybridisation - Radioactive DNA probes having repeated base sequences
complementary to possible VNTRs are poured over the nylon membrane. Due to its
natural affinity some of them will bind to the single stranded VNTRs and form double
stranded structure. This double stranded DNA structure remains embedded in the nylon
membrane. The nylon membrane is washed to remove extra probes.

Observation & Results:

Photography - An X-Ray film is exposed to the nylon membrane to mark the places where the
radioactive DNA probes have bound to the DNA fragments. The DNA bands due to radioactive
probe gives image on X-Ray film for documentation. These places are marked as dark bands
when X-Ray film is developed. This is known as Autoradiography.
The dark bands on X-Ray film represents DNA Fingerprints (=DNA Profiles).

Description & Importance:

The mutations in the genes produce alleles with different expressions, mutations in noncoding
repetitive DNA have no immediate impact. These mutations in the noncoding sequences have
piled up with time and form the basis of DNA Polymorphism (variations at genetic level arise due
to mutations). DNA Polymorphism is the basis of genetic mapping of human genome as well as
DNA Fingerprinting.
Short nucleotide repeats in the DNA are very specific in each individual and vary in number from
person to person but are inherited. These are the ‘Variable Number Tandem Repeats’
(VNTRs). These are also called minisatellites. VNTRs were first isolated and identified by A.
Wyman and R. White in 1980 at the University of Utah. Probability of having similar sets of
VNTRs in any two individuals is about 1 in 300 Million. Each individual inherits these repeats
from his/her parents which are used as genetic markers in a personal identity test. For Example,
a child might inherit a chromosome with six tandem repeats from the mother and the same
tandem repeated four times in the homologous chromosome inherited from the father. One half
of VNTR alleles of the child resemble that of the mother and other half with that of the father.
DNA profiling is a forensic technique in criminal investigations, comparing criminal suspects'
profiles to DNA evidence so as to assess the likelihood of their involvement in the crime. It is also
used in parentage testing, to establish immigration eligibility, and in genealogical and medical
research. DNA profiling has also been used in the study of animal and plant populations in the
fields of zoology, botany, and agriculture.

Significance & Application:

• Individuality- Like skin fingerprinting, DNA Fingerprinting can help to distinguish one human
being from another with exceptions of monozygotic twins.
• Paternity/Maternity Disputes- DNA Fingerprinting can identify the real genetic mother,
father and the offspring.
• Human Lineage- DNA from various probables is being studied to find out human lineage.
 • Hereditary Diseases- The technique is being used to identify genes connected with hereditary
diseases.
• Forensics- DNA Fingerprinting is very useful in the detection of crime and legal pursuits. DNA
Fingerprinting has proved that Dhanu, the human bomb, was the real murderer of Shri Rajiv
Gandhi, the former Prime Minister of India.
• Sociology- It can identify racial groups, their origin, historical migrations and invasions.
Genography is the study of migratory history of human species.

Reference:
Trueman’s Elementary Biology Volume II – K.N. Bhatia & M.P. Tyagi
Fundamentals Of Forensic DNA Typing – John M. Butler
DNA: The Story Of Genetic Evolution – James Watson
Wikipedia.org/wiki/DNA_profiling