DNA Profiling or Fingerprinting

Prepared by: Bersamina, Hanna Grace D.

Background:
developed by Professor Alec Jeffreys in 1984
It was first used in the law courts of England in 1987
to convict a man in a rape case.
It has now been used successfully in many crime and
paternity cases in worldwide.

 99.9% of human DNA sequences are the same in every

person
DNA profiling uses repetitive sequences that are highly
variable, called variable number tandem repeats
(VNTRs), particularly short tandem repeats
(STRs).
The analysis of variable number of tandem repeats
(VNTRs), to detect the degree of relatedness to
another sequence of oligonucleotides , making them
ideal for DNA fingerprinting.

AVariable Number Tandem Repeat (orVNTR) is a

location
in
agenomewhere
a
shortnucleotide
sequenceis organized as atandem repeat.
Minisatellite - aka Variable number of tandem repeats
(VNTRs)- usually composed of 9 to
80 bp or base
pairs.
Microsatellite aka Short tandem repeats (STR)
contains 2 to 5 nucleotides.

PROCESS:
1. Sample: sample of buccal swab,blood,saliva,semen, or
other appropriate fluid or tissue from personal items (e.g.
toothbrush, razor, etc.) or from stored samples (eg:
bankedspermorbiopsytissue
2. Processing of sample: only small amount of the sample
is used. E.g root of hair
3. Cutting - restriction enzymesare used to cut the DNA at
specific sites.
e.g an enzyme called EcoR1, found in bacteria, will cut
DNA only when the sequence 5..GAATTC..3 occurs.
4. Sizing and sorting - The DNA pieces are sorted according
to size by a agarose gel electrophoresis.

5. Transfer of DNA to nylon : The distribution of DNA pieces is

transferred to a nylon sheet by placing the sheet on the gel and
soaking them overnight.
6. Probing :Adding radioactive or colored probes to the nylon sheet
produces a pattern called the DNA fingerprint. Each probe typically
sticks in only one or two specific places on the nylon sheet.
7. DNA fingerprint : The final DNA fingerprint is built by using several
probes (5-10 or more) simultaneously. It resembles the bar codes used
by grocery store scanners.

Other methods:
RFLP (restriction fragment length polymorphism)
PCR analysis (Polymerase Chain Reaction)

RFLP (restriction fragment length

polymorphism)
RFLP analyzes the length of the
strands of the DNA molecules with
repeating base pair patterns.
The basic technique for detecting
RFLPs involves fragmenting a sample
of DNA by arestriction enzyme,
The resulting DNA fragments are then
separated by length through a process
known asagarose gel electrophoresis,
and transferred to a membrane via the
Southern blotprocedure.
Hybridizationof the membrane to a
labeledDNA probethen determines
the length of the fragments which
arecomplementaryto the probe.

PCR (Polymerase chain

reaction)
In this process, the DNA sample is
denatured
into
the
separate
individual strands.
Specific DNA primers are used to
hybridize to two corresponding
nearby sites on opposite DNA
strands in such a fashion that the
normal enzymatic extension of the
active terminal of each primer
(that is, the 3 end) leads toward
the other primer.
In this fashion, two new copies of
the sequence of interest are
generated.

Applications:
1. Diagnosis of inherited disorders :
Cystic fibrosis, hemophilia, Huntington's disease,
familial Alzheimer's, sickle cell anemia, thalassemia, and
many others.
2. Biological Evidence to Identify Criminals
3. Paternity disputes :
The use of DNA fingerprints in the court system is to
establish paternity in custody and child support litigation
4. Personal Identification

RED CELL GENOTYPING

Over the past 20 years the molecular

bases of almost all the major blood group
antigens have been determined. This
research has enabled development of DNAbased methods for determining blood group
genotype. The most notable application of
these DNA-based methods has been for
determining fetal blood group in
pregnancies when the fetus is at risk for
hemolytic disease of the fetus and
newborn.

Genetic basis of blood groups

Alleles that code for blood group polymorphism
Structure of gene products
A committee of ISBT( International Society of Blood
Transfusion) is responsible for the nomenclature of red
cell surface antigen and genes

Molecular Basis of blood group polymorphism

According to Blood Group Antigen Gene Mutation
Database (dbRBC)- There are 30 blood group system, 39
genes, 1 165 alleles.
ABO, P, Lewis, H, I ,MNS comprised of antigens that
are glycolipids
The rest of blood group system - comprised of antigens
that are direct consequences of amino acid variation in
the protein sequences