DNA

FINGERPRINTING
AND GEL
ELECTROPHORESIS
Noynay, Rigil Kent
Guyha, Chuck Brail
Villarin, Jemerie Dale
Dinglasa, Ericka Minette
Jumamil, Krisshia Mhae
Linis, Shanyne
Urbiztondo, Jianne Jahdiel

DNA fingerprinting
or DNA profiling
- any of several similar techniques
for analysing and comparing DNA
- 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, have exactly the
same DNA sequence, and those
segments will be statistically
unique.

Fingerprint

- very unlikely that any two

people would have exactly the
same DNA
information.

 The

test is used to determine

whether a family relationship
exists between two people, to
identify organisms causing a
disease, and to solve crimes.

 The

procedure for creating a

DNA fingerprintconsists of first
obtaining a sample of cells, such
as skin, hair, or blood cells,
which contain DNA. The DNA is
extracted from the cells and
purified.

Polymorphism

polymorphism is a clinically
harmless DNA variation that
does not affect the phenotype.
At the molecular level,
polymorphism is a variation in
nucleotide sequence from one
individual to another.

Polymorphisms

often occur in
the intervening sequences that
do not code for proteins. [Note:
Only a few percent of the human
genome actually encodes
proteins.]
All the common blood types,
such as the ABO blood group
system , are genetic
polymorphisms.

Tandem repeats

Polymorphism

in chromosomal
DNA can arise from the
presence of a variable number
of tandem repeats .
These are short sequences of
DNA at scattered locations in
the genome, repeated in
tandem (one after another).

 The

number of these repeat units

varies from person to person, but is
unique for any given individual and,
therefore, serves as a molecular
fingerprint.

Cleavage

by restriction enzymes
yields fragments that vary in length
depending on how many repeated
segments are contained in the
fragment.

Variation

in the number of tandem

repeats can lead to polymorphisms.

Single base changes

in DNA
About
90% of human genome
variation comes in the form of singlenucleotide polymorphisms, (SNPs,
pronounced snips), that is,
variations that involve just one
base .
The

alteration of one or more

nucleotides at a restriction site can
render the site unrecognizable by a
particular restriction endonuclease.
A new restriction site can also be

 In

either case, cleavage with an

endonuclease results in fragments
of lengths differing from the
normal, which can be detected by
DNA hybridization.

The

altered restriction site can be

either at the site of a diseasecausing mutation or at a site some
distance from the mutation.

Example

: Sickle cell anaemia.

Application of
DNA
Fingerprintin
g
)

Uses of DNA Profiling

DNA

profiling is
used to solve
crimes and
medical
problems

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).

Biological materials
used for DNA profiling
Blood
Hair
Saliva
Semen
Body

tissue/cells
Vaginal cells transferred to
the outside of a condom
during sexual intercourse.

DNA Profiling 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 (NB:it does not prove
they committed the crime).
If the profile doesnt 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 scene?
Suspects
Profile

Blood sample from

crime scene

Victims
profile

Solving Medical
DNA profiles Problems
can be used to determine
whether a particular person is the
parent of a child.
A childs 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 it is possible
to
identify DNA fragments in the
child
which are absent from the mother
and
must therefore have been

Is this man the father of the

child?
Mother

Child

Man

Famous cases
In

2002
Elizabeth
Hurley used
DNA profiling
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.

Famous Cases
O.J.

Simpson was
cleared of a double
murder charge in 1994
which relied heavily on
DNA evidence.

This

case highlighted
lab difficulties.

SUBSTAGES OF
DNA
FINGERPRINTING

1: A cell sample is taken- usually a

cheek
swab or blood test
2: DNA is extracted from sample
3: Cleavage of DNA by restriction
enzymethe DNA is broken into
small fragments
4: Small fragments are amplified by
the
Polymerase Chain Reactionresults in
many more fragments

5: DNA fragments are separated

by
electrophoresis
6: The fragments are transferred
to an
agar plate
7: On the Agar Plate specific DNA
fragments are bound to a
radioactive
DNA probe

8: The Agar Plate is washed free of

excess probe
9: An x-ray film is used to detect a
radioactive pattern
10: The DNA is compared to other
DNA samples

Why is DNA
Fingerprinting is done?
DNA fingerprinting is done to:
Find

out who a person's parents or

siblings are.

This

test also may be used to identify

the parents of babies who were
switched at birth.

Solve

crimes (forensic science)

Identify

a body

How is DNA
Fingerprinting is Done?

DNA fingerprint - is kind of like a

regular
fingerprint.

You are born with it, it is unique to

you (unless you have an identical
twin!), and you can leave it behind
wherever you go.

 But

unlike a fingerprint from

your hand, your DNA
fingerprint can't be found by
just "dusting for prints" like
they do on detective shows.

To find a DNA fingerprint, a

scientist has to first take the
DNA out of the nucleus of a
cell.

The cell that is used to

get a DNA fingerprint can
be:
a

skin cell

hair root cell

cheek

cell that gets washed out of

your mouth in your spit.

 This

is because your unique DNA

is the same in all of your cells.

The

goal is to analyze the DNA

in a way that shows scientists
the tiny differences in the DNA
of different people.

In

the past, scientists used a

technique called RFLP
(Restriction Fragment Length
Polymorphism)..

 RFLP

analysis - needs lots of DNA

But

sometimes only a little is left

behind at a crime scene. So
scientists found a way to use less
DNA.

They

worked out a method called

microsatellite analysis. But scientists
want to find ways to use even less
DNA

 They

also want to find a way to speed

up the process.
There are so many samples waiting to
be tested that labs can't handle them
all.
The wave of the future is something
called "lab-on-a-chip."
Lab-on-a-chip - a credit card sized
machine that you could load a tiny
sample into on the spot.
- would use tiny tubes and
pumps to perform all
steps normally done
by hand by scientists.

Advantages of DNA Fingerprinting

1. It is an easy and painless method for the subject being
tested. It is less invasive then taking a blood sample
2. It is an affordable and reliable technique
3. It can be conducted in a relatively short amount of time
4. Anyone at any age can be tested with this method
without any major concerns
5. There is a large variety of uses such as in legal claims,
missing persons cases, identification for the military, and
paternity and prenatal testing
6. The technique has used since 1984, making it highly
developed and improved

Disadvantages of DNA Fingerprinting

1. The sample of DNA can easily be ruined during the

process of DNA fingerprinting, causing the sample to
become completely useless for testing

2. The process itself is complex and tedious, and can give

results that may be hard to interpret
3. The test needs to be run on multiple samples, a
numerousamount of timesforideal accuracy. Commonly,
labs run each test twice with four samples.
4. Privacy issues could occur if the information isn't kept
secure at the lab. Personal information legally can only be
released with a written order. This personal information if
leaked, could potentially complicate insurance processes,
health care and job prospects for an individual

Alec Jeffreys
Born :
Alec John
Jeffreys
January 9, 1950
Oxford,Oxfordsh
ire, England,
United Kingdom
Age : 65 Years
old

He

is a professor of genetics at
theUniversity of Leicester,and he
became an honoraryfreemanof the City
ofLeicesteron 26 November 1992.In
1994, he wasknightedfor services to
genetics.

Heis

a Britishgeneticist, who developed

techniques forDNA Fingerprintingand
DNAprofiling which are now used
worldwide inforensic scienceto assist
police detective work and to resolve
paternity and immigration disputes.

Gel Electrophoresis

- method for separation and

analysis of
macromolecules
(DNA, RNA and
Proteins)
and their fragments based on
their size and charge.
- used in clinical chemistry to
separate
proteins by charge
and/or size
- used in biochemistryand
molecular
biologyto
separate a mixed population

- method of separation of
nanoparticles
- uses a gel as an anticonvective
medium and/or sieving medium
during
electrophoresis ~ the movement
of a charged particle in an
electrical field.

Gels suppress the thermal

convection caused by application
of the electric field, and can also
act as a sieving medium, retarding
the passage of molecules; gels can
also simply serve to maintain the

 DNA Gel electrophoresis - usually performed for

analytical purposes, often after amplification of
DNA viaPCR
- may be used as a
preparative technique prior to use of other
methods such asmass spectometry ,
RFLP ,PCR ,cloning , DNA Sequencing ,
orSouthern blottingfor further characterization.
Electrophoresis - a process which enables the
sorting of molecules based on
size
Sieving when nuclear acid molecules are
separated by applying an electric field to move
the negatively charged molecules through a
matrix of agaroseor other substances.
Proteins are separated by charge in agarose

 Using an electric field, molecules (such as DNA)

can be made to move through a gel made of
agaror polyacrylamide.
The electric field consists of a negative charge at
one end which pushes the molecules through the
gel, and a positive charge at the other end that
pulls the molecules through the gel.
The molecules being sorted are dispensed into a
well in the gel material. The gel is placed in an
electrophoresis chamber, which is then
connected to a power source.
When the electric current is applied, the larger
molecules move more slowly through the gel
while the smaller molecules move faster. The
different sized molecules form distinct bands on

 When separatingproteinsor smallnucleic

acids(DNA, RNA oroligonucleotides) the gel is
usually composed of different concentrations of
acrylamideand a cross-linker, producing different
sized mesh networks of polyacrylamide.
When separating larger nucleic acids (greater
than a few hundredbases), the preferred matrix
is purified agarose.
In both cases, the gel forms a solid, yet porous
matrix.
Acrylamide - is aneurotoxin and must be handled
using appropriate safety precautions to avoid
poisoning.

Agarose - composed of long unbranched

chains of uncharged carbohydrate without
cross links resulting in a gel with large
pores allowing for the separation of
macromolecules and macromolecular
complexes.

If charges are not all uniform then,

the electrical field generated by the
electrophoresis procedure will affect
the species that have different
charges and therefore will attract
the species according to their
charges being the opposite.
Cathode - negatively charged
- positively charged species
will
migrate towards it.
Anode - positively charged
- negatively charged species
will
migrate towards it.

Types of Gel in Gel

Electrophoresis
Agarose

- It is made from the natural

polysaccharide polymers extracted
from seaweed.
- are easily cast and handled compared
to other matrices, because the gel
setting is a physical rather than
chemical change.

 Samples

are also easily recovered.

After the experiment is finished, the
resulting gel can be stored in a plastic
bag in a refrigerator.
- do not have a uniform pore size, but
are optimal for electrophoresis of
proteins that are larger than 200 kDa.

 Agarose

gel electrophoresis can also be

used for the separation of DNA
fragments ranging from 50base pairto
several megabases (millions of bases),
the largest of which require specialized
apparatus. The distance between DNA
bands of different lengths is influenced
by the percent agarose in the gel, with
higher percentages requiring longer
run times, sometimes days.

Instead

high percentage agarose gels

should be run with apulsed field
electrophoresis (PFE), orfield
inversion electrophoresis.

 Polyacrylamide
is

used for separating proteins ranging

in size from 5 to 2,000 kDa due to the
uniform pore size provided by the
polyacrylamide gel.

Pore

size is controlled by modulating

the concentrations of acrylamide and
bis-acrylamide powder used in creating
a gel. Care must be used when creating
this type of gel, as acrylamide is a
potent neurotoxin in its liquid and
powdered forms.

 TraditionalDNAsequencing

techniques
such asMaxam-Glibert or Sangermethods
used polyacrylamide gels to separate DNA
fragments differing by a single base-pair in
length so the sequence could be read.

Most

modern DNA separation methods now

use agarose gels, except for particularly
small DNA fragments. It is currently most
often used in the field ofimmunologyand
protein analysis, often used to separate
different proteins of isoforms of the same
protein into separate bands. These can be
transferred into a nitrocellulose
orPVDFmembrane to be probed with
antibodies and corresponding markers, such
as in awestern blot.

 Starch
Partiallyhydrolysedpotato

starch
makes for another non-toxic
medium for protein
electrophoresis. The gels are
slightly more opaque than
acrylamide or agarose. Nondenatured proteins can be
separated according to charge and
size. They are visualised using
Napthal Black or Amido Black
staining. Typical starch gel
concentrations are 5% to 10%.

Advantages of Gel
Electrophoresis
Improved Diagnosis
-Medical laboratories use electrophoresis to
diagnosis blood disorders.

Simplicity Electrophoresis
-is a fast and easy technique

Low-Cost Material
-The material necessary to perform
electrophoresis costs little and is easy to prepare

Reliability
-Of all the benefits electrophoresis offers,
reliability is the most essential

Disadvantages of Gel
Electrophoresis

Electrophoresis Has Limited Sample Analysis

- Electrophoresis is specific to whatever tissue
you've sampled.

Electrophoresis Measurements Are Not Precise

-Gel electrophoresis can effectively separate
similar proteins with different weight

Only Certain Molecules Can Be Visualized

-Electrophoresis is excellent at separating and
identifying medium- to large-sized biomolecules.

Thank You For

Listening!!!