DNA Typing methods
Objectives
• To explain the extraction and analysis of DNA from
sample retrieved from crime scene.
• To describe Qualitative and quantitative estimation of
DNA
Learning outcomes
• To make student understand with types of
DNA typing process
• To understand differnce between STR’s and
VNTR’s and their role in forensic analysis
WHAT IS DNA?
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FINGERPRINTING
CONVENTIONAL DNA
FINGERPRINT FINGERPRINT
WHAT IS DNA
FINGERPRINTING?
• A technique by which the DNA of an
individual can be compared with that found in
a sample or another individual.
• This technique shows similarities and
dissimilarities between the DNA present in
different individuals.
• SIR ALEC JEFFREY at the University of
Leicester developed DNA fingerprinting in the
mid 1980s.
SAMPLE
FORENSIC EVALUATION
ISOLATION OF DNA
QUANTIFICATION
ANALYSIS OF DNA TYPE
RFLP ANALYSIS AND/OR PCR ANALYSIS
INTERPRETATION
FLOW CHART FOR DNA FINGERPRINTING
HOW CAN WE EXTRACT THE DNA?
DNA EXTRACTION
INORGANIC ORGANIC CHELEX
EXTRACTION EXTRACTION EXTRACTION
QUANTIFICATION
• CHECK THE PURITY OF DNA.
HOW IS DNA
FINGERPRINTING DONE?
• RFLP:- Restriction • PCR:-
fragment length Polymerase chain
polymorphism focuses reaction, is used to
on segments that produce multiple
contain sequences of copies of segments
repeated DNA bases, from a very limited
which vary widely amount of DNA .
from person to person.
DISADVANTAGES OF RFLP
• More costly.
• Requires a large sample of fresh DNA.
• Takes longer than PCR.
PROCEDURE OF RFLP
VNTRs STRs
• Repeated sequences of • Having smaller
base pairs. These repeat units.These
sequences, called sequences, called
Variable Number
Short Tandem
Tandem Repeats
(VNTRs).
Repeats (STRs).
• Contain 20-100 base • Contain 2-7 base
pairs. pairs.
VNTR PATTERNS
GEL ELECTROPHORESIS
• Refers to electromotive
force.
• Agarose gel containing
sample is placed in this
buffer-filled box and
electrical current is
applied.
• Staining.
• Destaining.
• Visualization.
SOUTHERN BLOTTING
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MAKING A RADIOACTIVE PROBE
• Obtain some DNA
polymerase. Put the
DNA to be made
radioactive (radio
labeled) into a tube.
• Introduce nicks and add
individual nucleotides to
the nicked DNA.
• Add the DNA
polymerase.
• Nucleotides
present in the
strand are replaced
with radioactive
nucleotides.
• Splitting of DNA
strands.
• The radioactive
DNA, now called a
probe.
CREATING A HYBRIDIZATION
REACTION
• Binding of two
genetic sequences.
• Denaturation.
• Put the denatured
DNA, saline liquid
and the probe into
one bag.
• The probe will bind
to the denatured
DNA.
THE POLYMERASE CHAIN
REACTION (PCR)
• It is a biochemistry and
molecular biology
technique for
exponentially
amplifying DNA, via
enzymatic replication,
without using a living
organism.
Thermal cycler for PCR
A STRIP OF EIGHT PCR TUBES
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PROCEDURE:-
ADVANTAGES OF PCR
• Takes less time.
• Less costly.
• Requires only a small sample.
• Produces multiple copies.
• Can be performed on older samples.
APPLICATIONS
• Paternity testing • Personal
identification.
• Criminal
identification
• Diagnosis of
inherited disorders .
Electrophoresis of
PCR amplified
DNA fragments
FAQ,S
• What is DNA? What is Fingerprinting and types of
fingerprinting.
• How Quantification of DNA is done.
• How is DNA fingerprinting done?
• Difference between STR and VNTR
• For forensic analysis Why PCr method is preffered
over RFLP.
References
• KS Reddy (2007):The essentials
of forensic medicine and toxicology, pg 212
• Richard Li (2008):Forensic Biology edition 2nd
Crc press publication pg.220
