Minhaj University Lahore

Academic Year 2023-2027

Department: Criminology and Forensic Science

Name of Assignment: Forensic DNA Typing

Submitted To: Prof. Aniza Batool

Full Name: Ahsan Ullah Tariq Hushmat

Roll no: 13.

Subject: Forensic Techniques And DNA Analysis Forensic Techniques & DNA Analysis
Date:10/August/24

DNA FINGERPRINTING:
Synonyms: DNA Typing, DNA Analysis, DNA Profiling, DNA Studies, Genetic Typing,
DNA Testing, Genetic Fingerprinting

DNA Fingerprinting is the technique applied to prepare a profile of an individual containing

DNA [ Deoxyribo Nucleic Acid] in their cell nucleus. More specifically this technique is applied
to human beings. Although DNA fingerprinting is also done in Bacteria, Fungus, plants etc. In
this chapter we are concerned with only Human DNA Fingerprinting.
In a very simple term DNA Fingerprinting is taking photograph or auto-radiograph of DNA
which is very similar to that of taking photograph of Human Being but since DNA is a very
small molecule so we need some sophisticated technique to do so that is DNA Fingerprinting.
The photograph/ auto-radiograph is taken in Ultraviolet light with specific DNA sequence
emitting radioactive waves by a radioactive Dye. The Photograph so produced is in the form
of Light and Dark Band and sequence of bands will tell us about the genetic make of a
particular repeated sequence.
DNA Fingerprinting is a technique employed to assist in the identification of individuals by
their respective DNA profiles. DNA profiling technique was first reported in 1985 by
Professor Alec Jeffreys at the University of Leicester in England
Introduction to DNA:
Deoxyribo-Nucleic acid DNA is a long chain of base pairs (Adenine, Thiamine, Cytosine and
Guanine) and two chains of base pairs joins with each other in a reverse pattern to form
complete DNA molecule. These double helical chains are attached on histone protein to form
a chromosome responsible for the characteristic features in living organism. Gene is a small
portion of this DNA responsible for a particular character or trait or may responsible to code
for a particular protein. Adenine and Thiamine connect with each other by two bond while
cytosine and Guanine bonded with each other with three bonds.
A-C-C-T-G-A-T-A-G-G-T-C-T-A-A
The opposite base pair strands will be
T-G-G-A-C-T-A-T-C-C-A-G-A-T-T
DNA strands are read in a particular direction from the top to bottom ( 5' or "five prime" end
to 3' or "three prime" end) as seen below:
5' A-C-C-T-G-A-T-A-G-G-T-C-T-A-A 3' 3' T-G-G-A-C-T-A-T-C-C-A-G-A-T-T 5'
The chemical structure of DNA is the same for all organism containing DNA. The difference
between organisms is the sequence of the base pairs.
Constituents of DNA
▪ Phosphates
▪ Deoxyribose Suger
▪ Organic Nitrogenous Base :
 Adenine
 Thymine
 Guanine
 Cytosine
▪ Single DNA molecule consist of 50-500 million base pairs
Chromosome: A rod-like structure of tightly coiled DNA, on histone proteins, found in the
cell nucleus of plants and animals. In Human Beings, there are 23 pairs of Chromosomes 22
pairs are common in both male and female 23rd pair is different for both sexes and is
responsible for the same.
Restriction enzyme: An enzyme which cuts DNA at specific sites always acts upon DNA in
the same manner.
Broadly the procedure included for DNA Fingerprinting is
▪ TAKE BIOLOGICAL SAMPLE
▪ EXTRACT WHOLE DNA
▪ CUT DNA IN SMALL PIECES
▪ RUN ON GEL
▪ TAKE PHOTOGRAPH AT DIFFERENT TIME
Methods of DNA Fingerprinting for forensic importance
Methods applied in DNA Fingerprinting:
1. RFLP Method or Restriction Fragments lengh polymorphism: In this method biological
samples like blood, semen, tissues, muscles, bones etc are collected and DNA is
extracted with the help of standard phenol-chloroform method or may be with the
help of commercially available DNA Extraction Kits. The Extracted DNA is mixed with
restriction enzymes which breaks the DNA into smaller pieces. The Fragmented DNA
is run on gel electrophoresis under the influence of electric field. The Smaller fragments
moves faster as compared to large fragments. The distribution of DNA band pattern
is transferred to a nylon sheet by placing the sheet on the gel and soaking them
overnight by a technique known as SOUTHERN BLOTTING . A radioactive DNA probe
is prepared which binds with specific DNA sequences to produce a fluorescent image.
It is known as hybridization. Hybridized probe fragments are visualized by auto-
 radiography. The bands on the X-Ray film appears corresponding to hybridized
fragments. This is DNA Fingerprint.
2. VNTR and STR: Variable number tandem repeats, represent specific locations on a
chromosome in which tandem repeats of 9-80 or more bases repeat a different number
of times between individuals. These regions of DNA are readily analyzed using the
RFLP approach and a probe specific to a VNTR locus. The fragments are a little shorter
than RFLPs (about 1-2 kilo base pairs), but are created through the exact same
process. Unlike VNTRs which analyze minisatellites that have repeat sequences of 9-
80 base pairs, STRs use microsatellites which have repeat sequences of only 2-5 base
pairs, introducing the “less is more” philosophy to the world of DNA fingerprinting.
This was a big step forward in forensic science since the length of DNA fragment being
analyzed is short enough to be amplified by polymerase chain reaction.
3. Polymerase Chain Reaction: PCR is done when the amount of recoverable DNA is small.
The Smaller amount of DNA is amplified with the process of PCR. It is like doing
photocopy of a small DNA fragments. In this method the two strands of DNA are
separated and allowed to regrow their own fragment under controlled tempeture and
availability of raw material [OLIGONUCLEOTIDE PRIMERS, DEOXYNUCLEOTIDE ] and
catalysts [Magnesium Chloride].

USES:IDENTIFICATION

▪ Mass Disaster cases including several casualties which need to be identified

as separate entity.
▪ Bomb Blast Cases
▪ Fragment Remains
▪ Unidentified bodies or Mutilated Remains
▪ Mass Casualties: Train and Air Accidents
▪ Doubtful Identification cases
▪ Missing Soldiers
▪ Missing Foreigners
▪ Wildlife identification
▪ Exhumation Cases

USES: PARENTHOOD
▪ Disputed Maternity
▪ Disputed Paternity
 ▪ Exchange of Child in hospital
▪ Exchange of child in early stage ▪ Founding Missing child after long
time.
▪ Transfer of Property cases.
▪ Citizenship Disputes
▪ Rape cases

USES: OTHERS
▪ Diagnosis of inherited disorders.
▪ Biological relationship for transplant
▪ Detection of Bacteria and other organisms.
▪ Hit and Run Cases of Road Traffic Accidents.
▪ Disputed Diagnosis in case of histopathological examination an doubtful
cases relating patient with particular sample.

Collection of Samples for DNA Extraction

▪ BLOOD
 Whole Liquid Blood: About 5ml of whole liquid blood is taken in preferably
EDTA vial.
 Blood Clot : whole blood clot is taken if size is small.
 Wet Blood Stains: Blood is taken on sterile cotton swab and air dried and then
sent to Lab in zip-locked polybags.
 Dried Blood Stains: whole dried blood is sealed in zip-locked polybag.
▪ SEMEN
 Dried seminal stain similarly as blood
 Wet seminal stains ▪ Hair Root.
▪ Saliva and Salivary Stains taken in glass tube ▪ Body tissue cells: taken in clean
sterile container.
▪ Fingernail Scraping taken in polybag.
▪ Postmortem samples like Muscle, Spleen, Bone marrow, organs, Vascular pulp of tooth
are taken in clean sterile plastic tube without any preservative. If delay is there then
freeze at – 20 C
Paternity Disputes
▪ VNTR patterns can be used to establish paternity and maternity. The patterns are so
specific that a parental VNTR pattern can be reconstructed even if only the children's
VNTR patterns are known
 ▪ Parent-child VNTR pattern analysis has been used to solve standard
fatheridentification cases as well as more complicated cases of confirming legal
nationality and, in instances of adoption, biological parenthood.

Advantages of DNA Fingerprinting

▪ DNA id stable molecule
▪ Small Quantity of Sample is needed ▪ Reliable technique ▪ Permanent record .
▪ Easy availability of DNA samples
▪ Repetition of same results with same or different samples.
▪ Lack of knowledge of Murdered/culprit.
Disadvantages
▪ Identical Twins
▪ Expensive and Sophisticated method
▪ Expert knowledge is needed
▪ Intermixing of samples of culprit with others available at crime scene.
▪ Lack of police training in sample collection.
▪ Require more time and hence accused can run away.
▪ Chances of error are there.
▪ Lack of availability of DNA Laboratories.
▪ Chances of deliberate samples exchanges.
▪ Decomposition of Samples adversely effect results

References:
1. Imran Sabri, Shazia Imran, Forensic DNA Isolation and Recovery in Blood and Semen.
Lamberts Academic Publisher, Germany. ISBN: 9783843381031.

2. Sabri I., Usmani J.A., Hanif S.A., Khan A.U.: Effect of temperature and ageing on
recovery of DNA in forensic blood samples. Indian Internet J. Forensic. Med. & Tox.
July- Sept. 2008; 6[3]:69-77.
 3. P V Guharaj, M R Chandran: Forensic Medicine, Second Edition. Orient Longman.
ISBN 139788125024880.
4. Kate Brinton, Kim-An Lieberman. Basics of DNA Fingerprinting:
[Link]
5. Reddy KSN: Essentials of Forensic Medicine and Toxicology: Publisher: K. Suguna
Devi.
6. Ballantyne, John, George Sensabaugh, and Jan Witkowski. DNA technology and
forensic science. Cold Spring Harbor, NY: Cold Spring Harbor Lab, 1989.
7. Jeffreys AJ, Wilson V, Thein [Link] 'minisatellite' regions in human
[Link]. 1985 Mar 7-13;314(6006):67-73.
8. Betsch, David (2005) DNA Fingerprinting in Human Health and Society.
[Link]
9. Christopher Lach, Thomas Patsis: DNA Fingerprinting: An Interactive Qualifying
Project Report [Link]
130417/unrestricted/[Link]

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