Biology

project

On
DNA Fingerprinting
 Index
Page
Topic no.
Certificate of authenticity 1
Acknowledgement 2
What is DNA fingerprint 3
Polymerase Chain Reactions(PCR) 4
VNTRs 5
Gel electrophoresis 6
Southern Blotting 9
DNA fingerprinting advantages 10
Forensic sciences 11
Famous cases solved by DNA
fingerprinting 12
Arbitrary 15
Reference 16
 Certificate of Authenticity 1

This is to certify that Richard Singh, T a student of

class 12th A has successfully completed the research
project on the topic DNA Fingerprinting under the
guidance of Mrs.Bindhu.
This project is absolutely genuine and does not
indulge in plagiarism of any kind. The references
taken in making this project have been declared at
the end of this project.

Signature (subject teacher) Signature

(examiner)
Acknowledgement: 2

I feel proud to present my project in biology on the

DNA Fingerprinting.
This project would not have been feasible without the
proper rigorous guidance of biology teacher
Mrs.Bindhu who guided me throughout this project in
every possible way. An investigatory project involves
various difficult lab experiments, which have to obtain
the observations and conclude the reports on a
meaningful note. These experiments are very critical
and in the case of failure may result in disastrous
consequences.
Thereby, I would like to thanks both Mrs.Bindhu and
lab assistant Mrs.Meena for guiding me on a
systematic basis and ensuring that in completed all
my experiments with ease.
Rigorous hard work has put in this project to ensure
that it proves to be the best. I hope that it proves to be
the best. I hope that this project will prove to be a
breeding ground for the next generation of students
and will guide them in every possible way.
 What is DNA fingerprinting? 3

DNA profiling (also called DNA fingerprinting, DNA testing, or

DNA typing) is a forensic technique used to identify
individuals by characteristics of their DNA. A DNA profile is a
small set of DNA variations that is very likely to be different in
all unrelated individuals, thereby being as unique to
individuals as are fingerprints (hence the alternative name
for the technique). DNA profiling should not be confused
with full genome sequencing. First developed and used in
1984, DNA profiling is used in, for example, parentage testing
and criminal investigation, to identify a person or to place a
person at a crime scene, techniques which are now
employed globally in forensic science to facilitate police
detective work and help clarify paternity and immigration
disputes. DNA fingerprinting has also been widely used in the
study of animal and floral populations and has
revolutionized the fields of zoology, botany, and agriculture.

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 ("identical") twins. DNA profiling uses
repetitive ("repeat") sequences that are highly variable,
called variable number tandem repeats (VNTRs), in
particular short tandem repeats (STRs), also known as
microsatellites, and mini satellites. VNTR loci are very similar
between closely related individuals, but are so variable that
unrelated individuals are extremely unlikely to have the
same VNTRs.
 Polymerase Chain Reaction (PCR) 4

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) 5

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
 Gel Electrophoresis 6

Fragments of DNA from restriction enzyme cleavage

are separated from each other when they migrate
through a support called an agarose gel
It is similar to the yummy food Jell-O gelatin
It is actually made out of some of the same
ingredients

The size-based separation of Molecules of DNA

separate based on size when an electric current is
applied to an agarose gel.

This method is called gel electrophoresis

 Gel Electrophoresis 7

The separated DNA fragments are then drawn out of

the gel using a nylon membrane

The nylon membrane is treated with chemicals that

break the hydrogen bonds in DNA and separate the
strands

The single stranded DNA is cross linked to the nylon

membrane.
By heat or UV light

Incubate the nylon membrane with a radioactive

probe of single stranded DNA complementary to the
VNTRs.
 Gel Electrophoresis 8

The radioactive probe shows up on photographic

film
Because as it decays it gives off light
The light leaves a dark spot on the film

Different individuals have different patterns of bands

These make up the fingerprint.

This Protocol is known as Southern Blotting.

Southern Blotting 9

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: 10
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 11

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.
Famous cases solved by DNA
fingerprinting 12

DNA Fingerprints, stray cigarette helped solve 1997 killing, sheriff

says
Amber Gail Creek, 14, was found dead in a Burlington marsh
Racine County Sheriff Christopher Schmaling on Tuesday identified
James P. Eaton as the 36-year-old Illinois man authorities suspect
killed Amber Gail Creek, a 14-year-old runaway girl from Illinois, in
1997. Credit: Racine County Sheriffs Department
The 1997 killing of a 14-year-old girl whose body was found in a Racine
County marsh has been solved 17 years later with old fingerprints and
new DNA evidence unknowingly provided by the alleged
killer, the sheriff said Tuesday. Racine County Sheriff 13
Christopher
Schmaling identified the suspect as James P. Eaton, a 36-year-old
Illinois man, and said he is being held on $1 million bail in the death of
Amber Gail Creek, an Illinois runaway.
"Over the last 17 years we have dedicated thousands of investigative
hours to bring this tragic and senseless murder to its resolution,"
Schmaling told reporters at a news conference. "Today that day is
here."
The break in the case stemmed from fingerprints left on the black
plastic bag used to suffocate the teen, Schmaling said, and
investigators were able to track Eaton and pull DNA evidence off one
of his cigarettes. Eaton, a Palatine, Ill., resident, faces charges in
Racine County of first-degree intentional homicide and hiding a
corpse. Schmaling, who did not take questions from reporters, said
Amber's family has asked for privacy. Amber ran away from a state-
operated juvenile shelter in Chicago on Jan. 23, 1997. She attended a
party at a motel in Rolling Meadows, Ill., the week of her death and was
last seen leaving the party and getting into a luxury car with a man.
On Feb. 9, 1997, two men stumbled upon her body while they were
searching for hunting sites at the Karcher Wildlife Refuge in the Town
of Burlington.
She had been sexually assaulted, and her body was found posed with
an upraised hand and the greeting "HI" written on the back of her
hand, according to Journal Sentinel archives. She was found with a $5
price tag on her arm from a bookstore in the Schaumburg, Ill., area.
Investigators spent more than a year and a half examining
more than 1,300 missing persons files before they identified 14
her, and the case drew national attention at the time, including a
profile on "America's Most Wanted" that aired Dec. 12, 1998.
Fingerprints taken from the plastic bag and DNA evidence from
Amber's body initially yielded no matches, despite the help of the FBI
and agencies around the country.
Then this year, through new fingerprint identification technology
used by a crime lab in Oklahoma, authorities were able to match the
prints found on the plastic bag to Eaton's prints, Schmaling said. Eaton
had been fingerprinted in Illinois for past, minor offenses, but his
name previously had not come up in the Amber Creek case.
After learning of the fingerprint match on Feb. 28, Racine County
investigators conducted surveillance on Eaton for several days, and
when he dropped a cigarette while waiting for a train in the Chicago
area, they picked it up for DNA evidence.
It matched the DNA found on Amber's body, Schmaling said."Our sense
of accomplishment is tempered by the pain and loss we know Amber's
family continues to confront every day," Schmaling said. "Our
thoughts and prayers remain with Amber's father, mother, other family
and loved ones."
It wasn't clear whether Amber and Eaton knew each other or how they
first would have come into contact. Eaton was 19 at the time of the
killing.
The sheriff said investigators are still looking for people who can
provide with information in the case. They are encouraged to contact
the Racine County sheriff's office at (262) 636-3225 or leave an
anonymous tip at Crime Stoppers of Racine at (888) 636-9330.
 Arbitrary 15

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 hybridisation of the DNA fragments using
a radio-labelled VNTR probe. Finally, the hybridised DNA
fragments are detected by a technique called
autoradiography conducted using an X-ray film.
Hybridisation 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, 16

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
www.bcps.org

http://www.csun.edu

www.nextgurukul.in

http://archive.jsonline.com/news/crime

https://en.wikipedia.org