Chapter Eleven – Forensic DNA Analysis

1. Chapter Eleven – Forensic DNA Analysis FRSC 8104 Criminalistics II Professor Bensley

2. FRSC 8104 – DNA Objectives • Understand the basic principles of DNA structure and analysis. • Recall the history and major steps in the development of Forensic DNA Analyses • Describe and detail the methods of DNA Extraction and Quantitation from an evidentiary sample. • Describe and detail the steps involved in the Polymerase Chain Reaction.

3. FRSC 8104 – DNA Objectives • Understand the importance of Short Tandem Repeats in Forensic DNA Analysis. • Understand the function of the CODIS system • Describe various techniques used in the visualization of amplified DNA. • Convey the important concepts of advanced DNA techniques to include Y-STR and Mitochondrial DNA analyses.

4. FRSC 8104 – DNA Outline • Introduction and Overview • Sir Alec Jeffreys (1985) • Colin Pitchfork • Results reported in DNA analysis

5. FRSC 8104 – DNA Outline • Biological and Genetic Basics – VIDEO! • Basic DNA Principles

6. Nucleotide Structure (dATP)

9. FRSC 8104 – DNA Outline • Chromosomes, Genes, DNA Markers

11. Locus (plural loci) is the specific location of a gene or DNA sequence on a chromosome. A variant of the DNA sequence at a given locus is called an allele. The ordered list of loci known for a particular genome is called a genetic map.

14. FRSC 8104 – DNA Outline • DNA Marker Nomenclature • Population Variation SEQUENCE POLYMORPHISM LENGTH POLYMORPHISM -------(AATG) (AATG) (AATG)-------- 3 repeats ------------(AATG) (AATG)------------ 2 repeats

15. FRSC 8104 – DNA Outline • Historical Methods • Pre 1985 • RFLP-Based DNA Typing RFLP Animation Southern Blot Animation

16. FRSC 8104 – DNA Outline • Early Based PCR Testing • HLA DQ Alpha / DQA1

17. FRSC 8104 – DNA Outline • Polymarker (PM+DQA1)

18. FRSC 8104 – DNA Outline • D1S80

19. FRSC 8104 – DNA Outline • Short Tandem Repeats

20. FRSC 8104 – DNA Outline • Silver vs. Fluorescent Staining Techniques

21. FRSC 8104 – DNA Outline • DNA Extraction • Organic Extraction • Chelex Extraction • FTA Paper • Solid Phase Extraction • Differential Extraction

22. FRSC 8104 – DNA Outline • DNA Quantitation • Why? • How? TAQMan

23. FRSC 8104 – DNA Outline • DNA Amplification (PCR) • Introduction • Why PCR? • Issues with PCR • PCR Components How PCR was Discovered What the?!?!?

24. FRSC 8104 – DNA Outline • The Process of Replication • DNA Polymerase • Okazaki Fragments DNA Replication Process Video

25. http://rst.gsfc.nasa.gov/Sect20/DNA-20replication.gif

27. FRSC 8104 – DNA Outline • Denaturation • Annealing • Extension • Multiplex PCR • PCR Inhibition Video of the PCR Process Another Video of the PCR Process

28. FRSC 8104 – DNA Outline • Short Tandem Repeat Markers (STRs) • Repetitious DNA • Microsatellites or STR

29. FRSC 8104 – DNA Outline • 13 CODIS STR Loci

30. The Thirteen CODIS STRs and Their Probability of Identities Source: The Future of Forensic DNA Testing: Predictions of the Research and Development Working Group. Washington, DC: National Institute of Justice. Department of Justice, 2000, p. 41.

31. FRSC 8104 – DNA Outline • DNA Separation and Detection • Electrophoresis • Slab Gels • Capillary CE Animation • DNA Detection

33. FRSC 8104 – DNA Outline • Advanced Techniques • Y-STR

34. FRSC 8104 – DNA Outline • Mitochondrial DNA