FORENSIC

GENETICS
DNA AND ITS APPLICATIONS IN FORENSIC
SCIENCES

Dr. Sadia Abdul Qayyum

Assistant professor Forensic Medicine
LNHMC
OBJECTIVES

• DNA molecular basis

• DNA Typing techniques (RFLP, PCR, STR, mtDNA, Y Chromosome
Analysis)
• DNA Evidence collection
• Uses of DNA in Forensic Sciences
USES OF DNA

1. To establish identity of a person

2. Identification in postmortem
practice
3. Paternity of the child is
determined
4. Evidence-guilty/not guilty
5. Organ transplantation
6. Human remains
7. Diseases
8. To identify sex.
9. To resolve disputes
DNA MOLECULAR BASIS

• Deoxyribonucleic acid, is a complex

molecule that contains all of the
information necessary to build and
maintain an organism.
• DNA has three types of chemical
component:
1. Phosphate
2. A sugar called deoxyribose
3. And four nitrogenous bases—adenine,
guanine, cytosine, and thymine.
• Two of the bases, adenine and guanine,
have a double-
ring structure characteristic of a type of
chemical called a purine
 The main questions that are asked to
address are:

1. Whose DNA is it?

2. From what body fluid has it
originated?
3. How did it get there?
4. Have the results been reported in a
fair and balanced way?
The following items may contain DNA
material:

1. Masks
2. Hats
3. Gloves
4. Clothing
5. Tools
6. Weapons
7. Sexual assault evidence kits
8. Underclothes
9. Bedding
10. Dirty laundry
11. Fingernail scrapings
12. Cigarettes
13. Toothpicks
14. Toothbrush
15. Facial tissue
16. Hairbrush
17. Eyeglasses
18. Condoms
19. Tape
20. Ligatures (rope, wire,
cords)
21. Stamps or envelopes
22. Cups/bottles
 DNA TYPING TECHNIQUES

• DNA typing is a laboratory procedure

that detects normal variations in a
sample of DNA.

1. Polymerase Chain Reaction (PCR)

Analysis.
2. Restriction Fragment Length
Polymorphism (RFLP)
3. Short Tandem Repeat (STR) Analysis.
4. Mitochondrial DNA Analysis.
5. Y-Chromosome Analysis.
• The DNA testing process is
comprised of four main steps,
including

Extraction
Quantitation
Amplification
Capillary electrophoresis
 RESTRICTION FRAGMENT LENGTH
POLYMORPHISM (RFLP)

• Technique developed to analyse variable

lengths of DNA fragments produced through
DNA digestion.
• It exploits variations in DNA sequences due
to the differing locations of restriction
enzyme sites.
• The method uses restriction endonucleases
to ‘digest’ the DNA by cutting it at specific
sequence patterns.
• The resulting restriction fragments are then
separated using gel electrophoresis and
transferred to a membrane using the
Southern Blot technique.
• After the separated DNA fragments are
transferred, probe hybridisation is used to
detect the fragments.
 POLYMERASE CHAIN REACTION (PCR)
ANALYSIS.

• Technique which allows for the

exponential amplification of DNA
fragments to lengths of approximately
10,000 base pairs.
• A single copy of a DNA fragment could
be amplified to millions of copies in
just a few hours.
• PCR is particularly beneficial in the
amplification of minute amounts or
degraded samples.
• In the first step of PCR, the two strands of the DNA double helix are physically
separated at a high temperature in a process called DNA melting

• In the second step, the temperature is lowered and the primers bind to the
complementary sequences of DNA.
APPLICATIONS

1. DNA cloning for sequencing, gene

cloning and manipulation, gene
mutagenesis
2. Functional analysis of gene
3. Diagnosis and monitoring of heredit
ary diseases, amplification of ancient
DNA
4. Analysis of genetic fingerprints
for DNA profiling and detection
of pathogens in infectious diseases
 SHORT TANDEM REPEAT(STR)
ANALYSIS

• This type of DNA analysis works to

examine individual areas in DNA.
• The differences for certain DNA
regions in one person versus another
can allow for distinguishing between
individuals.
• STR analysis involves the extraction of
nuclear DNA from cells in a sample
• Certain regions of the DNA that are
extracted are then amplified by the
polymerase chain reaction.
• Performs gel electrophoresis to find
out how many repeats of the STR
sequence exist.
• In criminal investigations, there
are thirteen regions that are
analysed and compared to
establish profiles.
• DNA databases used at the
government level involve the
sequence of these thirteen
regions.
• The chances of two people having
the exact same thirteen regions is
virtually impossible - likely one in
a billion.
 MITOCHONDRIAL DNA ANALYSIS

• Mitochondrial DNA (mtDNA) is

located outside the chromosomes of
a cell in the mitochondria.
• Children inherit mtDNA only from
their mother.
• An analysis of the mitochondrial
DNA (mtDNA) enables you to
discover your own family history in
the maternal line.
• Mitochondrial DNA offers
several benefits over nuclear DNA
when determining phylogenetic
pathways
• Maternal inheritance – mtDNA is
inherited from the mother only,
providing a much more direct genetic
lineage
IMPORTANCE

• Inherited changes in mitochondrial

DNA can cause problems with
growth, development, and function
of the body's systems
• Samples that involve very old bones,
teeth or hair shafts are simply not
feasible to analyse through
techniques such as STR or RFLP
analysis
• Mitochondrial DNA (mtDNA) has
provided forensic scientists with a
valuable tool for determining the
source of DNA recovered from
damaged, degraded, or very small
biological samples
 METHOD OF DNA EVIDENCE
COLLECTION

• Collection: of stained item itself.

• Scraping: of dried stain
• Tape lifting: of dried stain
• Swabbing: of stain
• The most common places to
find DNA are in blood, semen,
hair, and saliva.
• You can also find DNA in urine,
skin cells, perspiration, teeth,
bone, and internal organs, so
the need to examine the scene
carefully.
• In situations where you are
collecting DNA, the key thing is
to guard against
contamination.
• Use disposable tools, such as
disposable tweezers, to
handle evidence.
• Never place any evidence
that may contain DNA in
plastic bags
• Biological evidence must be
stored dry and cold
• In laboratory, DNA samples
may be stored at 4°C or -
20°C; long term at -70°C
 FORENSIC DNA ANALYSIS IN
PAKISTAN

• Punjab Forensic Science Agency (PFSA), being the largest forensic

DNA laboratory in Pakistan, has been helping in solving rape, murder,
dead body identification, sibship, and parentage cases.
• Having started its operations since 2012, PFSA has received
approximately 44,417 cases for DNA analysis out of which 36,954
cases have been solved successfully while approximately 7463 cases
are pending for several reasons (DNA and Serology Department)
https://link.springer.com/article/10.1186/s41935-019-0162-2
https://sahsol.lums.edu.pk/law-journal/dna-evidence-pakistani-
courts-analysis
• Picture from Google images.
• Read from text book of parikh and krishan Vij.

THANK YOU