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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
1
Arbitrary 5
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 side
during 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

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