AISSCE (2017-18)

INVESTIGATORY PROJECT ON
DNA FINGER PRINTING

SUBMITTED BY:-
Name: Sitakanta Rout
Roll No:
Under the guidance of Mrs Sandhya Rani
Pradhan
 Bona-fide

This is certify that Sitakanta Rout has

Successfully completed the investigatory project
DNA finger printing under the guidance
Of Mrs. Sandhyarani Prandhan.

Signature of Teacher: Signature of Examiner:

 ACKNOWLEDGEMENT

I express my gratitude to my Biology teacher Mrs. SANDHYARANI

PRADHAN for her guidance. I consider it has a great privilege to
have the opportunity to present this work under her expert
supervision. I am deeply indebted to her for her guidance and
encouragement.
I am also thankful to all my faculties of biology department
Mrs. Sandhyarani Pradhan
Mr. B C Nayak (Laboratory Assistant) for their cooperation.

Sitakanta Rout
Class: XII
 Bio-data

Name : Sitakanta Rout

Class: XII
Roll No:
School: Buxi Jagabandhu English
Medium school.
Project:DNA Finger printing
Guiding Teacher:Mrs. Sandhya
Pradhan
Examination: AISSCE
Academic year: 2017-2018
 INDEX

1.Bona-fide

2. Acknowledgement.

3. Bio-data.

4. Introduction.

5. History of fingerprinting.

6. What is DNA fingerprinting?

7. Biological material used for DNA fingerprinting.

8. Methods of Collection.

9. Polymerase Chain reaction (PCR).

10. VNTRs.

11.SSR and STR.

12. Methods of DNA fingerprinting.

13. DNA fingerprinting Applications.

14. Southern Blotting.

15. DNA fingerprinting advantages.

16. Forensic Science

17. Arbitrary

18. Reference
 INTRODUCTION
The process of DNA fingerprinting was invented by Alec Jeffreys at the
University of Leicester in 1985. He was knighted in 1994.

DNA fingerprinting or DNA profiling, any of several similar techniques

for analyzing and comparing DNA from separate sources, used especially in
law enforcement to identify suspects from hair, blood, semen, or other
biological materials found at the scene of a violent crime. It depends on
the fact that no two people, save identical twins, have exactly the same
DNA sequence, and that although only limited segments of a person's DNA
are scrutinized in the procedure, those segments will be statistically
unique.
 History of DNA Fingerprinting
Up to 1984, the only method of establishing and authenticating
personal identification was by the fingerprint process.

DNA fingerprinting technique was devised in 1985 by Alec Jeffrey at

University of Leicester in England, while working on the sequences
within myoglobin gene.

What is DNA
Fingerprinting?
It is a technique, by which an individual can be identified at molecular
level. With the advancement of science and technology STR analysis
has become very popular in forensic laboratories. Scientists have
chosen repeating sequences in the DNA, which are present in all
individuals on different chromosomes, and are known to vary from
individual to individual except in identical twins. These are used as
genetic markers to identify the Individual.
 Biological Material Used for DNA
fingerprinting
COMMON BIOLOGICAL SAMPLES ENCOUNTERED

Whole Fresh Blood

Blood Stain
Seminal Stain
Hard Tissues (Bones)
Soft Tissues (body organs)
Hair

METHODS OF COLLECTION:

Whole blood Sample: Sterile needle should be used while withdrawing or

collecting blood.

Blood stain: Should be picked up preferably on sterile cotton gauge using sterile
forceps and blade.

Seminal stain: Should not be touched by hand especially the stain portion. Should
be picked up with sterile forceps.

Hard Tissues: Bones-- bones should be picked up using gloves, Kept at a place
where there are no chances of environmental contamination. It should be allowed
to dry completely.

Soft Tissues: Body organs should be collected using forces and wearing gloves. It
should be kept in a sterile container.

Hair: Hair roots are preferred for the analysis. Hair roots should be picked up
using sterile forceps.
PRESERVATION:

Whole Blood: Blood should be collected in sterile container containing an

anticoagulant. The mostly preferred is EDTA. It should be mixed properly but
gently for some time. The container should be covered with parafilm to avoid
slippage. Should be kept it at 40C or using ice during transportation till it reaches
laboratory for analysis.

Blood stain: Blood stain should be dried properly. In semi dry stain there, is a
possibility of bacterial growth thus chances of having contamination. Drying
should be avoided using electric fans. After complete drying it should be wrapped
in a fresh blotting paper and packed in a Zip lock poly bag. No preservative is
required. It can be transported at environmental temperature.

Seminal stain: Likewise seminal stain should also be dried properly. In semi dry
stain there, is a possibility of bacterial growth thus chances of having
contamination. It should not be dried using electric fans. After drying it should
be wrapped in a fresh blotting paper and packed in a Zip lock poly bag. No
preservative is required. It can be transported at environmental temperature.

Hard Tissue: No preservative is required. The hard tissues should be wrapped in

the blotting paper and placed in a zip lock poly bag.

Soft tissue: It should be placed at 40C or in Ice till it reaches laboratory for
analysis.

Hair: Hair roots should be placed in a blotting paper and then packed in a zip lock
poly bag. It requires no preservative and can be transported at environmental
temperature.

FORWARDING:

After packing all these zip lock poly bags, they should be numbered and sealed.
Each sample, even from the same case should be sealed separately. The seal has
to be from the office of the forwarding authority. The covering letter should
bear the signature of the forwarding authority and a copy of seal should be
enclosed. Wherever photographs are to be attached, the forwarding authority or
Doctor collecting blood sample should attest them. While collecting fresh blood,
there should be the signatures of minimum two (2) witnesses.

For forwarding other biological samples also, the signature of minimum two
witnesses is required.
Polymerase Chain Reaction (PCR)
If there is only a small amount of DNA available for DNA
Fingerprinting the amount of DNA is increased by using
a technique called PCR. PCR is a method of DNA replication in a test tube.
Like All DNA Polymerases Taq polymerase can only add
To the 3 end of an existing nucleotide
A DNA primer that is complementary to the template is used to supply that
3 end
 VNTRs (variable number tandem repeats)
After we isolate the DNA and amplify it with PCR

We then treat the DNA with restriction enzymes

cut DNA at specific sequences

Everyones DNA is different, so everyones DNA will cut

at different sites

This results in different sized fragments

The different sized fragments are called restriction

fragment length
polymorphisms, or RFLPs

We can observe the

different sized fragments
in an experiment that
separates DNA based on
fragment size called Gel
Electrophoresis

Everyone has genetic

sequences called
variable number tandem
repeats, or
VNTRs
 Everyone has different amounts of
VNTRs

The VNTRs make the different sized

RFLPs

VNTRS (variable number of tandems)

1. VNTRs or minisatellites surrounded by

conserved restriction sites.
2. A small DNA sequence is arranged
tandemly in many copy numbers.
3. The copy no varies from
chromosome to chromosome in an
individual.
4. The no. of repeats shows very high
degree of polymorphism.
5. As a result the size of the VNTR varies
from 0.1 to 20kb.
 SSRs ( Single Sequence Repeats) Or
STRs ( Short Tandem Repeats) Or
Microsatellites.

1. These sequences normally do

not code for any proteins.
2. They form a large portion of the
human genome.

3. These sequences show high

degree of polymorphism and
form the basis of DNA finger
printing.

4. Polymorphism refers to the

variation at genetic level.

5. Since DNA from every tissue such

as blood, hair follicle, skin, bone,
saliva, sperm etc) from an
individual shows the same
degree of polymorphism they
became useful tool in
identification in forensic
applications.
 The Steps of DNA Fingerprinting
DNA fingerprinting involves a number of intensive and important steps in order to
fully complete and develop and DNA fingerprint of a father, a suspect or a person
involved in an immigration problem.

1. The process of DNA fingerprinting starts with isolating DNA from any part
of the body such as blood, semen, vaginal fluids, hair roots, teeth, bones,
etc.

2. Polymerase chain reaction (PCR) is the next step in the process. In many
situations, there is only a small amount of DNA available for DNA
fingerprinting. Because of this, in a test tube, DNA replication is must occur
to make more DNA. The DNA and the cells will undergo DNA replication in
order to make more DNA to be tested.

3. After the DNA is isolated and more copies of the DNA have been made, the
DNA will be tested. The scientist will treat DNA with restriction enzymes
(an enzymes that cuts DNA near specific recognition nucleotide sequences
known as restriction sites).
-This will produce different sized fragments which are known as restriction
fragment length polymorphisms (RFLPs).
-These fragments can then be observed doing an experiment called gel
electrophoresis which separates DNA based on fragment sizes.

4. Gel electrophoresis is the next step in this process of DNA fingerprinting.

During gel electrophoresis, an electrical current is applied to a gel mixture,
which includes the samples of the DNA.
-The electric current causes the DNA strands to move through the gel. This
separates the molecules of different sizes.
-The fragments of separated DNA are sieved out of the gel using a nylon
membrane (treated with chemicals that allow for it to break the hydrogen
bonds of DNA so there are sing strands).
5. The DNA (single stranded) is cross-linked against the nylon using heat or a
UV light.

6. The probe shows up on photographic film because the strands of DNA decay
and give off light. In the end it leaves dark spots on the film which are also
known as the DNA bands of a person. What make up the fingerprint are the
unique patterns of bands. The pattern of bands are different because we
are all different and unique (other than identical twins).

7. Once the filter is exposed to the x-ray film, the radioactive DNA sequences
are shown and can be seen with the naked eye. This creates a banding
pattern or what we know as DNA fingerprints. This technique is called
southern blotting.

These fingerprints can be used to

determine which hair strand belongs to
which person for example.
DNA fingerprints of children should be
similar to the their parents fingerprints,
although they may not be the same. Some
bands will match one parent and other
bands can match the other parent. With
the bands of both of those parents, they
make the bands and the identity of the
child.
Below is an illustration of how DNA fingerprinting is done and the
process a scientist must go through to find the patterns of DNA.
DNA fingerprinting: Methods
A common procedure for DNA fingerprinting is restriction fragment length
polymorphism (RFLP). In this method,DNA is extracted from a sample and
cut into segments using special restriction enzymes RFLP focuses on contain
sequences of repeated DNA bases, which vary widely from person to person.
The segments are separated using a laboratory technique called
electrophoresis, which sorts the fragments by length. The segments are
radioactively tagged to produce a visual pattern known as an autoradiograph,
or DNA fingerprint, on X-ray film. A newer method known as short tandem
repeats (STR) analyzes DNA segments for the number of repeats at 13
specific DNA sites. The chance of misidentification in this procedure is one in
several billion. Yet another process, polymerase chain reaction , is used to
produce multiple copies of segments from a very limited amount of DNA (as
little as 50 molecules), enabling a DNA fingerprint to be made from a single
hair. Once a sufficient sample has been produced, the pattern of the alleles
(see genetics ) from a limited number of genes is compared with the pattern
from the reference sample. A no match is conclusive, but the technique
provides less certainty when a match occurs.
 DNA fingerprinting: Applications

DNA fingerprinting can be applied in the following scenarios:

Establishment of paternity and Maternity

Establishment of the parentage for child swapping cases
Establish the identity of the rapist in rape cases
Identification of mutilated remains In murder, bomb blast, air crashes
etc.
Wild life identification, and
Seed authentication.
Southern Blotting
A Southern blot is a method used in molecular biology for
detection
of a specific DNA sequence in DNA samples. Southern blotting
combines transfer of electrophoresis-separated
DNA fragments to a filter membrane and subsequent
fragment detection by probe hybridization.
DNA Fingerprinting advantages:
DNA fingerprints can be used to determine which
bone fragments belong to which individual.
DNA fingerprints of children should be similar to the those
of parents. DNA fingerprinting can show which individuals are
the parents of specific children.
 Forensic science
Forensic science is the application of science to criminal and
civil laws, mainlyon the criminal sideduring criminal
investigation, as governed
by the legal standards of
admissible evidence and
criminal procedure.

Forensic scientists collect,

preserve, and analyze
scientific evidence during
the course of an investigation. While some forensic scientists
travel to the scene of the crime to collect the evidence
themselves, others occupy a laboratory role, performing
analysis on objects brought to them by other individuals.[1]

In addition to their laboratory role, forensic scientists testify

as expert witnesses in both criminal and civil cases and can work
for either the prosecution or the defence. While any field could
technically be forensic, certain sections have developed over
time to encompass the majority of forensically related cases.
 Arbitrary
It is the small difference in base pair sequences of DNA that make the
phenotypic appearance of each individual unique. An easier and quicker solution to
comparing DNA sequences is DNA fingerprinting. In human beings, ninety-nine per
cent of DNA base sequences are identical and are known as the bulk genomic DNA.
The remaining one per cent DNA base sequences differ and are present as a small
stretch of repeated sequences known as repetitive DNA. DNA fingerprinting
identifies the differences in this region. To separate both genomic as well as
repetitive DNA the process of density gradient centrifugation is carried out. As
satellite DNA is lighter and bulk DNA is heavier, so they get separated on the
basis of their density. Graphical representation shows bulk genomic DNA as a
major peak and repetitive DNA as smaller peaks known as satellite DNA.
Satellite DNA is highly repetitive and consists of non-coding sequences. Based on
the length of the segment, base composition and number of repetitive units
satellite DNA can be classified as mini-satellite DNA and micro-satellite DNA.
Mini-satellite is a section of DNA which has a variable number of tandem repeats
or VNTR. This step is followed by the hybridization of the DNA fragments using a
radio-labeled VNTR probe. Finally, the hybridized DNA fragments are detected
by a technique called autoradiography conducted using an X-ray film.
Hybridization with the VNTR probe results in an autoradiogram, which produces
several bands of different sizes. These bands provide a characteristic pattern to
an individuals DNA and vary from one individual to another except in identical or
monozygotic twins. Today, the accuracy of the DNA fingerprinting technique has
further improved due to the advent of the polymerase chain reaction or PCR,
where multiple copies of a single DNA sequence can be made. DNA polymorphism
is the guiding principle behind genetic mapping and therefore it helps in the DNA
fingerprinting technique. The DNA fingerprinting technique was developed by Alec
Jeffreys. DNA fingerprinting technique helps in crime investigation, paternity
testing, determining genetic and population diversity and studying evolution and
speciation.
 References

NCERT CLASS 12th TEXTBOOK.

AAKASH MATERIAL CLASS 12th
http://www.csun.edu
www.nextgurukul.in
https://en.wikipedia.org
SCHOOL LIBARAY.