DNA ANALYSIS

MODULE 14
WHAT IS DNA?
WHAT IS DNA?
▪ Deoxyribose Nucleic Acid or Deoxyribonucleic Acid- is chemical
substance found in all cells whose composition has been passed on
from parent to their children.

WHAT DOES DNA DO?

▪ DNA contains genetic information
▪ DNA code contains instructions needed to make proteins & molecules
essential for growth, health & development.

DNA since 1986

WHAT IS DNA?
Some common proteins are:

▪ HEMOGLOBIN-carries oxygen from lungs to cells

▪ INSULIN-regulates metabolism
▪ KERATIN- protein found in hair
WHAT DOES DNA LOOK LIKE?
▪ DOUBLE HELIX – like a twisted ladder
DNA STRUCTURE
▪ The DNA ladder is made up of building blocks called
NUCLEOTIDES.

▪ What is a nucleotide?
DNA BASES

BASES are Nitrogen

containing
molecules.
DNA BASES

▪ Bases pair up to form the rungs of the ladder.

▪ A (Adenine) pairs with T (Thymine)
▪ C (Cytosine) pairs with G (Guanine)
FACTS ABOUT DNA Only one side of
the ladder is
written
▪ DNA is written as the sequence of aforementioned bases

▪ In humans, there are 3.3 billion base pairs in the DNA within each cell.
▪ Approximately 99.9% of the sequence of these 3.3 billion base pairs is identical for
all humans and performs the same function. Example of these functions include the
existence in each person of two eyes, head, hair and other biological functions
common to humans.
▪ 0.1% of difference is more than enough to distinguish us from one another.
▪ No 2 people have identical DNA sequence except from identical twins.
WHERE IS DNA?
▪ DNA is found in the cells of our body.
WHERE IS DNA?
▪ The DNA sequence, or order of the base pairs, is the same for every cell in the
body, with the exception of reproductive cells (egg & sperm cells). Each cells
contain only one-half of the person’s DNA.
SOME CELLS IMPORTANT TO FORENSIC SCIENCE ARE:
WHERE IS DNA?
▪ DNA in the nucleus is packed into the chromosomes.
WHERE IS DNA?
BIOLOGICAL EVIDENCE THAT
CAN BE SUBMITTED FOR DNA
ANALYSIS
▪ Blood and bloodstains
▪ Semen and seminal stains
▪ Hairs with follicle or root
▪ Salive or buccal swab
▪ Bones & Organs
▪ Tissues & Cells
CASES WHERE DNA CAN BE
OF HELP
▪ Sexual Assault
▪ Murder
▪ Homicide
▪ Robbery
▪ Hit and Run
▪ Extortion
▪ Paternity Case
▪ Identification of remains from mass disaster cases and missing persons
HOW DOES DNA
DIFFER AMONG
HUMANS?
HOW DNA DIFFER AMONG
HUMANS?
▪ There are several ways the sequence can be different:
DNA ANALYSIS
2 MAIN TYPES OF DNA
PROFILING

▪ RESTRICITION FRAGMENT LENGTH POLYMORPHISMS

▪ SHORT TANDEM REPEATS

RESTRICTION FRAGMENT
LENGTH POLYMORPHISM
(RFLP)
▪ A technique in which organisms may be differentiated by analysis of patterns
derived from cleavage of their DNA.

▪ If two organisms differ in the distance between sites of cleavage of a particular

restriction endonuclease, the length of the fragments produced will differ when the
DNA is digested with a restriction enzyme.
RESTRICTION FRAGMENT
LENGTH POLYMORPHISM
(RFLP)
▪ Take the pattern GCGC and imagine it occurs more than once in the DNA. The
number of times it occurs is unique to the individual. The restriction enzyme chops
the DNA in two at every place where the GCGC pattern occurs.
RESTRICTION NUCLEASE
▪ are enzymes that recognize a specific DNA sequence, called a restriction site, and
cleave the DNA within or adjacent to that site.

▪ For example, the restriction endonuclease EcoR I, isolated from the

bacterium Escherichia coli, recognizes the following sequence:
5′GAATTC3′
3′CTTAAG5′
A restriction enzyme is present to cleave the DNA strand with the following pattern:
5’GAATTC3’
3’CTTAAG5’
Determine what DNA fragments will be produced for each individual.
RFLP PROCEDURE
RFLP PROCEDURE
RFLP PROCEDURE
Gel electrophoresis:
• The restriction fragments have negative charge and can be separated by a
technique called GEL ELECTROPHORESIS, which separates the pieces of DNA
based on their size.
INTERPRET THE RESULT OF
DNA FINGERPRINTING
INTERPRET THE RESULT OF
DNA FINGERPRINTING
INTERPRET THE RESULT OF
DNA FINGERPRINTING
INTERPRET THE RESULT OF
DNA FINGERPRINTING
INTERPRET THE RESULT OF
DNA FINGERPRINTING
SHORT TANDEM REPITITION
ANALYSIS
SHORT TANDEM REPEATS
▪ The most common type of DNA profiling today for criminal cases and other types of
forensic use
▪ Examines how often base pairs repeat in specific loci, or locations, on a DNA
strand.
▪ As the name implies, short tandem repeat (STR) analysis is a method of
determining an individual's DNA profile by counting the number of times a small
DNA sequence (short tandem repeat unit) is repeated at a specific chromosomal
location. STR analysis consists of three processes: amplification, electrophoresis,
and interpretation.
▪ The number of repeats in each stretch is inherited and is easily detected
SHORT TANDEM REPITITION
ANALYSIS
STR
ANALYSIS
STEPS
PCR AMPLIFICATION

▪ During the PCR reaction, STRs are amplified using primer sequences binding to the
STR flanking regions.
POLYMERASE CHAIN
REACTION –DNA
AMPLIFICATION
CAPILLARY
ELECTROPHORESIS
Electrophoregram-analy
zed and translates result
into graph
INTERPRETATION OF RESULT
INTERPRETATION OF RESULT
INTERPRETATION OF RESULTS
MITOCHONDRIAL
DNA
MITOCHONDRIAL DNA
▪ Sometimes, a sample can be old and will no
longer have nuclear material in the cell, which
poses a problem for the other types of DNA
analysis. With mitochondrial DNA analysis,
however, mitochondrial DNA can be removed,
thus having important ramifications for cases that
were not solved over many years.

▪ Inherited from the mother only

MITOCHONDRIAL DNA
Advantages:
▪ More sensitive (less DNA needed), degrades slower
than nuclear DNA
▪ Can be used in cases where nuclear DNA cannot
(hair without root, skeletal remains)

Disadvantages:
▪ All people of same maternal line will be
indistinguishable (less discriminatory)
▪ More work, more time consuming, more costly