Chapter 7 DNA Fingerprinting By the end of

Chapter 7 DNA Fingerprinting By the end of this chapter you will be able to: o o 1 Explain how crime scene evidence is collected and processed to obtain DNA Describe how radioactive probes are used in DNA fingerprinting Explain how DNA evidence is compared for matching Explain how to use DNA fingerprinting to identify DNA from a parent, child, other relative, or a nonrelated individual

Dr. Alec Jeffrey’s 2

History of Biological Evidence in Forensics o o o 3 DNA fingerprinting • Also known as DNA profiling • Used with a high degree of accuracy • No 2 people have the same DNA except identical twins Biological evidence is examined for the presence of inherited traits EX. Skin, blood, saliva, urine, semen, hair Some forensics laboratory techniques were originally developed for other purposes. Can you name any? Medical diagnosis, treatments, inherited traits, blood typing

DNA o 4 No two people have the same DNA, with the exception of identical twins Karyotyping – the analysis of chromosomes determines abnormal chromosomes Normal Downs Syndrome

The Function and Structure of DNA o o o 5 DNA contains the genetic material of a cell Chromosomes are located in the cell nucleus Chromosomes contain long DNA strands wrapped around proteins

The Function and Structure of DNA o o o 6 Double Helix – found by Watson and Crick 1953 (Nobel Peace Prize) Chromosome is made up of two strands of DNA tightly coiled around protein molecules Can be teased apart by dissolving the histone sleeve that holds the DNA strand together

The Function and Structure of DNA o o Nitrogenous Bases-pairs of molecules that form the rungs of the DNA “ladder” Four types of Bases • • 7 A (adenine) C (cytosine) G (guanine) T (thymine)

DNA 8

The Function and Structure of DNA o Base-Pairing Rules • • • 9 What would pair with CGTCTA? adenine (A) binds only with thymine(T) Cytosine (C) binds only with guanine (G) 46 Chromosomes (23 pairs) in nucleus of human cells

DNA • • Nuclear DNA – In the chromosomes Mitochondrial DNA - is circular and only inherited from the mother Male = XY Mother = XX *each only give one chromosome. Therefore, who determines the sex of the offspring? Father, because he can give a Y chromosome 10

The Function and Structure of DNA o o 11 DNA sequences (CGTCTA) that determine our inheritance are genes. (give instructions) When genes come from different parents for the same trait they are referred to as alleles. EX. Mother gives allele or a gene for normal hemoglobin, Father gives allele for a gene for abnormal hemoglobin.

The Function and Structure of DNA Human GENOME The total amount of DNA in a cell o Contains 1. Chromosomal DNA – 3 billion base pairs that are responsible 12 for making proteins exons (encoded DNA) & introns (unencoded DNA) 23, 688 encoded genes (exons), each gene containing 3, 000 base pairs - this is only 1. 5 percent of the DNA in the genome 2. Mitochondrial DNA – 98. 5 percent of the DNA (introns) non-protein producers “junk DNA”

Human Genome Project Begun formally in 1990, the U. S. Human Genome Project was a 13 -year effort coordinated by the U. S. Department of Energy and the National Institutes of Health. The project originally was planned to last 15 years, but rapid technological advances accelerated the completion date to 2003. Project goals were to o identify all the approximately 20, 000 -25, 000 genes in human DNA, o determine the sequences of the 3 billion chemical base pairs that make up human DNA, o store this information in databases, o improve tools for data analysis, o transfer related technologies to the private sector, and o address the ethical, legal, and social issues (ELSI) that may arise from the project. Human Genome Project 13

Human Genome Project Gene-Splicing 14

DNA Identification o o 15 o Polymorphisms: Non-coded DNA that contain unique patterns of repeated base sequences that are unique to individuals DNA Fingerprinting (Profiling) isolates and analyzes polymorphisms DNA fingerprints appear as a pattern of bands on X-ray film.

DNA Profile Two types of repeating DNA sequences Variable Numbers of Tandem Repeats (VNTR) o The number of repeats varies from person to person 9 to 80 bases in length • • Short Tandem Repeats (STR) o 2 to 5 bases in length Shorter lengths make STRs easier to use than VNTRs • • o VNTR and STR data are analyzed for • • 16 tissue matching inheritance matching

DNA Profile Matching o Tissue Matching • o Inheritance Matching • 17 Two samples that have the same band pattern are from the same person Each band in a child’s DNA fingerprint must be present in at least one parent

DNA Population Databases o Population genetics: • • o Examples • 18 the study of gene variations among groups of individuals, used to calculate the % of people who have a particular allele. Analyze the probability of a random person with the same alternative form of a gene (an allele) • Identifying the suspect in a crime Identifying an alleged father in a paternity case

Sources of DNA o o 19 Biological evidence: examples include saliva, blood, skin, hair, seminal fluid Individual evidence is capable of identifying a specific person. Small amounts may be only trace evidence Polymerase chain reaction (PCR) technique generates multiple copies of DNA evidence

Collection and Preservation of DNA 1. Use disposable gloves and collection 2. 3. 4. 5. 20 instruments Avoid physical contact, talking, sneezing, and coughing in the evidence area Air-dry evidence and put it into new paper bags or envelopes Dry or freeze the evidence Keep evidence cool and dry during transportation and storage

Preparing DNA Samples for Fingerprinting Extraction • DNA is mixed with special enzymes Restriction • Enzymes cut apart the DNA in specific places forming different sized fragments Amplification • DNA is separated within an agarose gel Electrophoresis • 21 An electric current is passed through the gel separating the fragments by size

Preparing DNA Samples for Fingerprinting Extraction 1. Cells are isolated from biological evidence such as blood, saliva, urine, semen, and hair 2. The cells are disrupted to release the DNA from proteins and other cell components 3. The DNA can be extracted from the cell nucleus 22

DNA Preparation Restriction Ø DNA is cut by restrictive enzymes. ØThese enzymes recognize a unique pattern of DNA bases and will cut at that specific location ØThe length of these DNA bases may vary 23

Preparing DNA Samples for Fingerprinting Amplification o o 24 VNTR analyses—polymerase chain reaction (PCR) can be used to amplify the DNA that contains the VNTRs STR profiles—restriction enzymes are unnecessary; PCR allows the amplification of the strands with STR sequences

Electrophoresis 25

Electrophoresis 26

Preparing DNA Samples for Fingerprinting Electrophoresis o o o 27 The method of separating the molecules within an electric field based on the size of the DNA fragments DNA samples are placed in gels through which electronic currents are passed DNA fragments line up in bands along the length of each gel

Electrophoresis o o 28 An electrophoresis apparatus running five sample of DNA Arrows show the movement of the negatively charged DNA fragments through the gel matrix

Probes o DNA probes • • • o 29 identify the unique sequences in a person’s DNA by using radioactive probes are made up of different synthetic sequences of DNA bases complimentary to the DNA strand bind to complimentary bases in the strand (see the fragmentary DNA bands above) In most criminal cases, 6 to 8 probes are used

Analysis of DNA Fingerprints and Applications Bands and widths are significant in matching samples of DNA 30 Paternity

DNA Database o o o 31 United States uses the “Combined DNA Index System” (CODIS) Similar to the “Automated Fingerprint Identification System” (AFIS) Rape, murders and child abuse require DNA samples to be stored on CODIS.

Analysis of DNA Fingerprints and Applications o DNA fingerprinting can • match crime scene DNA with a suspect • determine maternity, paternity, or match to another relative • eliminate a suspect • free a falsely imprisoned individual • identify human remains 32

. . . Summary. . o o 33 DNA contains the information needed for replication in a sequence of nitrogenous bases. DNA analysis allows even a small sample of tissue to be identified with a single individual. DNA contains, in non-coding regions called junk DNA, many repeated sequences that vary in number between individuals. These differences between individuals can be used to produce a DNA fingerprint for an individual.

. . . . Summary o o 34 Polymerase chain reaction (PCR) for DNA amplification has largely eliminated the problem resulting from the tiny samples usually available. DNA evidence must be collected carefully to avoid contamination with other DNA analysis involves extraction, electrophoresis, and visualization. DNA profiles are kept by police agencies in electronic databases.