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Copyright © 2009 Pearson Education, Inc.
Application of Biotechnology in DNA Fingerprinting and Forensic Analysis
Copyright © 2009 Pearson Education, Inc.
Introduction to DNA Fingerprinting and Forensics
• Forensic science – intersection of law and science
• Historic examples
• Early 1900s – fingerprints
• DNA Fingerprinting – 1985
• Unique signature found in each person’s genetic
Copyright © 2009 Pearson Education, Inc.
What Is a DNA Fingerprint?
• DNA fingerprints is a way of identifying a specific individual,. It is also known as genetic fingerprinting or DNA profiling.
• Uses: identifying paternity or maternity and for identifying criminals or victims
• Every individual carries a unique set of genes
• Chemical structure of DNA is same, but the order of the base pairs differs
• Every cell contains a complete set of DNA that identifies the organism as a whole
• Only one tenth of 1% of DNA differs from person to person
Copyright © 2009 Pearson Education, Inc.
Fingerprinting • DNA fingerprinting uses a specific type of DNA sequence,
known as a microsatellite, to make identification
• Microsatellites are short pieces of DNA (1-6 base) which repeat many times (1-100 times) in a given person's DNA.
• In a given area, microsatellites tend to be highly variable, making them ideal for DNA fingerprinting. By comparing a number of microsatellites in a given area, one can identify a person relatively easily.
• Also called short tandem repeats (STR)
• FBI has chosen 13 unique STRs for testing
• Combined DNA Index System (CODIS)
Copyright © 2009 Pearson Education, Inc.
Paternity/maternity verification
• The sections of DNA used in DNA fingerprinting, although highly variable, are passed down from parents to their children
• no child has pairs that their parents do not have
Copyright © 2009 Pearson Education, Inc.
Forensic
• forensics is considered a very critical part of any investigation
• Witnesses can often be mistaken, have fading memories, or even pass away, forensics "tells the same story" no matter how many times it is tested, or how many years have passed.
• DNA fingerprinting is very attractive because it doesn't require actual fingerprints
• All the DNA sections are contained in every cell, any piece of a person's body, from a strand of hair to a drop of blood, may be used to identify them using DNA fingerprinting
Copyright © 2009 Pearson Education, Inc.
• Two Main Types of Forensic DNA Testing
• RFLP (restriction fragment length polymorphism)
• Requires larger amounts of DNA
• DNA cannot be degraded
• PCR (polymerase chain reaction)
• Less DNA and DNA can be partially degraded
• Extremely sensitive to contaminating DNA
Copyright © 2009 Pearson Education, Inc.
What Is a DNA Fingerprint?
DNA profiling depends on a small portion of the genome
• Exons code proteins
• Introns do not code for proteins
DNA fingerprint use the introns for identification the individu
Copyright © 2009 Pearson Education, Inc.
Prokaryotes vs eukaryotes
The introns contain repeated
sequences of between 1 and 100
base pairs
Called variable number tandem
repeats (VNTR’s)
Some VNTR’s are inherited
from mother and some from
father
Copyright © 2009 Pearson Education, Inc.
Preparing a DNA Fingerprint
• DNA Collection
• Search for sources of DNA
• Collection requires attention :
• Wear disposable gloves; change them frequently
• Use disposable instruments
• Avoid talking, sneezing, and coughing
• Avoid touching any item that might contain DNA(face, nose, or mouth)
Copyright © 2009 Pearson Education, Inc.
Preparing a DNA Fingerprint
Enemies of Evidence
• Sunlight and high temperature
• Bacteria
• Moisture
DNA fingerprinting is a comparative process
• Samples from crime scene must be compared to suspect DNA
• Best sample from suspect DNA is fresh, whole blood
Copyright © 2009 Pearson Education, Inc.
RFLP
• A molecular method of genetic analysis that allows individuals to be identified based on unique patterns of restriction enzyme cutting in specific regions of DNA
• The technique takes advantage of the polymorphisms in individual pe ople's genetic codes.
• Polymorphism: multiple forms of a single gene that can exist in an individual or among a group of individuals.
Copyright © 2009 Pearson Education, Inc.
Preparing a DNA Fingerprint
• RFLP (restriction fragment length polymorphism) Analysis
• cutting a particular region of DNA with known variability, with restriction enzymes,
• Separating the DNA fragments by agarose gel electrophoresis and determining the number of fragments and relative sizes. The pattern of fragment sizes will differ for each individual tested.
RFLP focuses on segments that contain sequences of repeated DNA bases, which vary widely from person to person.
Copyright © 2009 Pearson Education, Inc.
Preparing a DNA Fingerprint
• Southern Blot Technique
• Transfer DNA fragments from gel to nitrocellulose or nylon membrane
• Membrane incubated with a probe • Short strand of complementary DNA with a radioactive or
fluorescent tag
• Targeted area on the DNA fragment is called a locus
• Expose X-ray (photo) film to membrane to obtain permanent record of results