DNA

FINGERPRINTING
By:Parth Shah
Prinal Khimasia
Shrey Shah
Vikash Makadya
M. PHARM ( Pharmacology)
NMIMS UNIVERSITY, MUMBAI.

How do you figure out that someones

DNA is more similar to anothers?
The primary method of assessing
similarities is by use of DNA fingerprinting
or DNA restriction analysis.
This process makes use of special
proteins called restriction enzymes and
sections of the chromosome called
tandem repeats

Definition
Technology using tandem repeats of
individuals to identify individuals is
known as DNA fingerprinting

DNA Fingerprinting
By Taylor Cindric

DNA Fingerprinting is a method where:

a persons genetic traits, genes, are used to
make specific strings of DNA letters that are
cut into patterns of shorter strings separated
by length these banding patterns can
identify a unique human being!

Basis of genetic fingerprinting:

Most of our DNA is identical to each other, however,
there are inherited regions of our DNA that can
vary from person to person (such variations are
termed as polymorphisms). One such class of
polymorphisms is known as tandem repeats, which
vary within the individual of the species. This
forms the basis of genetic fingerprinting.

Tandem repeats
In a Eukaryotic genome, Tandem repeats are an
array of consecutive repeats, known as
satellites.
They are of three types based on migration
when centrifuged in CsCl density gradient.
1. Satellites.
2. Minisatellites.
3. Microsatellites.

Satellite DNA
Illustration of satellite bands.
By using buoyant density
gradient centrifugation, DNA
fragments with significantly
different base compositions
may be separated, and then
monitored by the absorption
spectra of ultraviolet light.
The main band represents the
bulk DNA, and the "satellite"
bands originate from tandem
repeats.

SATELLITES
The size of a satellite DNA ranges from
100 kb to over 1 Mb.Other satellites
have a shorter repeat unit. Most
satellites in humans or in other
organisms
are
located
at
the
centromere.

Minisatellites
The size of a minisatellite ranges from 1 kb to
20 kb. One type of minisatellites is called
variable number of tandem repeats (VNTR).
Its repeat unit ranges from 9 bp - 80 bp. They
are located in non-coding regions. The number
of repeats for a given minisatellite differs
between individuals. This feature is the basis
of DNA fingerprinting.

Microsatellites
Microsatellites are also known as short
tandem repeats (STR), because a repeat unit
consists of only 1-6 bp and the whole
repetitive region spans less than 150 bp.
Similar to minisatellites, the number of
repeats for a given microsatellite may differ
between
individuals.
Therefore,
microsatellites can also be used for DNA
fingerprinting.

Variable Number of Tandem Repeats

(VNTR):
repeats of 9 to 80 base pairs (bp), total length is 500
to 23,000 bp, very specific due to length and repeats,
testing is expensive and time-consuming, degrade in
older DNA samples due to random breaking of DNA
strands

Amplified Fragment Length

Polymorphisms (AmpFLP):
repeats of 8 to 16 bp, total length 100 to 1300 bp,
shorter and less susceptible to degradation, first loci to
be used in forensic analysis

Short Tandem Repeats:

repeats of 2 to 7 bases, total length 100 to 400 bp,
shorter yet thereby less susceptible to breakage,
these loci are the current standard in forensic
laboratory analysis, ideal size for PCR amplification

Single Nucleotide Polymorphisms (SNP):

a single base change as a result of mutation, not
commonly useful to forensic investigators, can be
potentially used to distinguish identical twins

Restriction Fragment Length

Polymorphisms (RFLP)
The
term
Restriction
Fragment
Length
Polymorphism, or RFLP refers to a difference
between two or more samples of homologous DNA
molecules arising from differing locations of
restriction sites, and to a related laboratory
technique by which these segments can be
distinguished.
.

 Commonly pronounced rif-lip.

Its analysis was the first DNA profiling technique
cheap enough to see widespread application.
It is an important tool in genome mapping.
Localization of genes for genetic disorders.
Determination of risk for disease, and paternity
testing

 A restriction enzyme cuts the DNA molecules at every

occurrence of a particular sequence, called restriction
site.
For example, HindII enzyme cuts at GTGCAC or GTTAAC.
If we apply a restriction enzyme on DNA, it is cut at every
occurrence of the restriction site into a million restriction
Any mutation of a single nucleotide may destroy or create
fragments each a few thousands nucleotides long.
the site(CTGCAC or CTTAAC for HindII) and alter the
length of the corresponding fragment.
The term polymorphism refers to the slight differences
between individuals, in base pair sequences of common
genes.

RFLP ANALYSIS TECHNIQUE:

RFLP analysis is the detection of the change in the
length of the restriction fragments.
The basic technique for detecting RFLPs involves
fragmenting a sample of DNA by a restriction enzyme,
which can recognize and cut DNA wherever a specific
short sequence occurs, in a process known as a
restriction digestion.
The resulting DNA fragments are then separated by
length through a process known as agarose gel
electrophoresis.
Then transferred to a membrane via the Southern blot
procedure

Hybridization of the membrane to a labeled DNA probe

then determines the length of the fragments which are
complementary to the probe.
Each fragment length is considered an allele, and can be
used in genetic analysis.

Stages of DNA Profiling

Stage 1:
Cells are broken down
to release DNA.
If only a small amount
of DNA is available it can
be amplified using the
polymerase chain reaction
(PCR).

Stages of DNA Profiling

Step 2:
The DNA is cut into fragments using restriction
enzymes.
Each restriction enzyme cuts DNA at a specific base
sequence.

Stages of DNA Profiling

The sections of DNA that are cut out are called
restriction fragments.
This yields thousands of restriction fragments of all
different sizes because the base sequences being cut
may be far apart (long fragment) or close together
(short fragment).

Stages of DNA Profiling

Stage 3:
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.

Stages of DNA Profiling

A radioactive material is
added which combines with
the DNA fragments to
produce a fluorescent
image.
A photographic copy of the
DNA bands

Stages of DNA Profiling

Stage 4:
The pattern of fragment distribution is then
analysed.

Biological materials used for DNA

profiling
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.

APPLICATIONS
1.Paternity and Maternity
person inherits his or her VNTRs from his or her
parents .
Parent-child VNTR pattern analysis has been used
to solve standard father-identification cases
2.Personal Identification
The notion of using DNA fingerprints as a sort of genetic
bar code to identify individuals.

3.

Criminal Identification and Forensics

DNA isolated from blood, hair,
skin cells, or other genetic
evidence left at the scene of a
crime can be compared
FBI and police labs around
the U.S. have begun to use
DNA fingerprints to link
suspects
to biological evidence
blood or semen stains, hair,
or items of clothing

4.Diagnosis of Inherited Disorders

diagnose inherited disorders in both
prenatal and newborn babies
These disorders may include cystic
fibrosis, hemophilia, Huntington's disease,
familial Alzheimer's, sickle cell anemia,
thalassemia, and many others.
5.Developing Cures for Inherited Disorders
By studying the DNA fingerprints of relatives who
have a history of some particular disorder
identify DNA patterns associated with the disease

Advantages of DNA Fingerprinting

1.Unsurpassed discriminatory potential :
Complete blood group testing allows discrimiation of one
person in several thousand and HLA typing one in several
million
DNA typing can routinely provide exclusion probabilities on
the order of one in billions

2.Exquisite sensitivity
DNA can be amplified
smaller sample sizes are adequate
allows rather small samples to be split and submitted
for testing to more than one laboratory
3.Application to any body tissue
DNA testing can be conducted with any sample having
nucleated cells
For example hairs, semen, urine and saliva

4.DNA is stable in comparison to proteins

resistant to degradation by common environmental
insults
DNA is also long-lived in comparison to protein

Disadvantages of DNA Fingerprinting

1. Problems with determining probability
.
.
.
.
.

DNA fingerprinting is not 100% assured

VNTR are results of genetic inheritance
not distributed evenly across all populations
cannot have a stable probability of occurrence.
Due to allele frequencies in different population or
ethnics groups, the probability of match can range
from 1 in 20 to 1 in 2 billion.

 Occurrence of certain VNTR pattern depends on an

individuals genetic background.
Big problem in determining the VNTR patterns of
heterogeneous genetic composition of interracial
individuals
For example, the frequency of a specific allele may
be 4% in Asians instead of 1% as it is in Northern
Europeans.
Contamiination of the sample
Shifting of bands produces wrong information.

THANK YOU