DNA Fingerprinting

CLASS:III B.Sc.
, MICROBIOLOGY
(VI SEMESTER)
Presented by
Dr. K.Anandhi M.Sc., SET., M.Phil., Ph.D.,
Head,Department of Microbiology
Shrimati Indira Gandhi College
Trichy-2
SYNOPSIS
 DNA Fingerprinting definition
 Invention
 Samples used for DNA Fingerprinting
 Steps in DNA Fingerprinting
 Uses of DNA Fingerprinting with Example
 Questions
What is DNA
fingerprinting ?
 DNA fingerprinting is a way to
identify a certain individual, rather
than simply identifying a particular
species or a particular trait.
 A technique used scientists to
distinguish between individuals of the
same species using only samples of
their DNA.
Who invented it ?
The process of DNA
fingerprinting was invented
by Alex Jeffreys in 1985
Biological samples used for
DNA fingerprinting
 Blood
 Hair
 Saliva
 Semen
 Body tissue cells
 DNA samples have
been obtained from
vaginal cells transferred
to the outside of a
condom during sexual
intercourse.
Steps in DNA fingerprinting
There are 8 steps for
DNA Fingerprinting
 Step 1: Isolation of DNA
 DNA must be
recovered from cells
or tissue. Only a
small amount of
blood, hair, or skin is
needed to isolate DNA
Step 2:Digestion of DNA by restriction
endonucleases
 The DNA is cut into fragments using
restriction enzymes.
 Each restriction enzyme cuts DNA at a
specific base sequence.
 The sections of DNA that are cut out are called
restriction fragments.
 This yields thousands of restriction fragments of all

differt sizes because the base sequences being cut
may be far apart (long fragment) or close together
(short fragment).
 Commonly used RE are (hae III, Hinf I, Alu I etc.)
 Reaction mixture is incubated overnight at 37.C

 Step 3: Electrophoretic
separation of different fragments
Fragments are separated
on the basis of size using
a process called gel
electrophoresis.
DNA fragments are
injected into wells and an
electric current is applied
along the gel.
 DNA is negatively
charged so it is attracted
to the positive end of the gel.
 The shorter DNA
fragments move faster
than the longer fragments.
DNA is separated on
basis of size
Step 4: Transfer DNA on
Nylon/Nitrocellulose membrane
 The DNA fragments are
transferred to a nylon sheet
by placing the sheet on the
gel and soaking them overnight
by the process southern blot.
 Step5: Probing/probe
labeling
 Adding radioactive or
colored probes to the
nylon sheet which is
complementary to
target sequences.
 Each probe only
sticks to one or two
specific places on the
sheet.
Step 6: Hybridization
labeled probe DNA should be
hybridized with the
complementary sequences
located on nylon membrane for
the detection of position of later
Membrane is washed to remove
non specific binding and
clearing of the background
 Step 7: Autoradiography
 To detect the sequences in genome
bound with the hybridized
radioactive probe on membrane
Technique involves alignment of
hybridized membrane with X-ray
film
 The X-ray film alter its
development shows multiple no. of
bands that looks like bar codes and
known as DNA fingerprints.
 Step 8: Interpretation
of band patterns
Analysis of band
patterns of different
individuals
comparison of
position of bands
 Computer software
are also available for
for the analysis of
DNA fingerprints
Uses of DNA fingerprinting
 Diagnosis of Inherited
Disorders
 Helps diagnose disorders in
both prenatal and newborn
babies
 Disorders may include
cystic fibrosis, hemophilia,
Huntington’s disease,
familial Alzheimer’s, sickle
cell anemia, thalassemia,
and much more.
 Crime
 Forensic science is the use
of scientific knowledge in
legal situations.
 The DNA profile of each
individual is highly
specific.
 The chances of two
people having exactly the
same DNA profile is
30,000 million to 1
(except for identical
twins).
DNA fingerprinting can solve crimes
 The pattern of the DNA profile is then compared

with those of the victim and the suspect.
 If the profile matches the suspect it provides strong
evidence that the suspect was present at the crime
scene (Note: it does not prove he committed the crime).
 If the profile doesn’t match the suspect then that
suspect may be eliminated from the enquiry.
Example
A violent murder occurred.
 The forensics team retrieved a blood sample from the

crime scene.
 They prepared DNA profiles of the blood sample, the

victim and a suspect as follows:
Was the suspect at the crime ?
Solving Medical Problems
DNA profiles can be used to determine whether a
particular person is the parent of a child.
A child’s paternity (father) and maternity(mother) can
be determined.
This information can be used in Paternity suits
Inheritance cases
Immigration cases
Example : A Paternity TEST
By comparing the DNA profile of a mother
and her child
 identify DNA fragments in the child which

are absent from the mother and must
therefore have been inherited from the
biological father.
Is this man the father of the
child?
Personal Identification
The U.S. armed services
are just beginning a
program where they collect
DNA fingerprints from all
personnel for later use,
incase they need to identify
casualties or missing
people.
 Some other uses of DNA fingerprinting
 Identification of carcass of tissues
Detection of somatic mutations or cancer Pathogen
identification
Detection of loci controlling quantitative traits or
disease resistance
 Sex determination
 Individual identification
Famous cases
In 2002 Elizabeth
Hurley used DNA
Fingerprinting to prove
that Steve Bing was the
father of her child
Damien
Famous cases
 Colin Pitchfork was the
first criminal caught
based on DNA
fingerprinting evidence.
He was arrested in 1986
for the rape and murder
of two girls and was
sentenced in 1988.
QUESTIONS
2MARKS
1.What is DNA Fingerprinting
2.Who invented DNA Fingerprinting
3.Write the uses of DNA Fingerprinting
5MARKS
4. Explain about the applications of DNA Fingerprinting
10 MARKS
5. Describe in detail about DNA Fingerprinting ,Steps
involved and the uses of it.

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CLASS:III B.Sc.

 This yields thousands of restriction fragments of all

 Commonly used RE are (hae III, Hinf I, Alu I etc.)

 Reaction mixture is incubated overnight at 37.C

 The pattern of the DNA profile is then compared

 The forensics team retrieved a blood sample from the

 They prepared DNA profiles of the blood sample, the

 identify DNA fragments in the child which

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