Biology

project
 On
DNA
Fingerprinting

Index
Page

Topic no.

1
€
 Certificate of authenticity

Acknowledgement 2
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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 “Munendra” 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 thank Mrs.Nilima
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 HYPERLINK
"https://en.wikipedia.org/wiki/Minisatellite" HYPERLINK
"https://en.wikipedia.org/wiki/Minisatellite" HYPERLINK
"https://en.wikipedia.org/wiki/Minisatellite" HYPERLINK
"https://en.wikipedia.org/wiki/Minisatellite" HYPERLINK
"https://en.wikipedia.org/wiki/Minisatellite" HYPERLINK
"https://en.wikipedia.org/wiki/Minisatellite"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

 – Everyone’s DNA is different, so everyone’s 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,
mainly—on 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
Sheriff’s 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 individual’s
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 Jeffrey’s. 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