Download FRSC 7104 - Chapter One Objectives

Chapter Eleven –
Forensic DNA Analysis
FRSC 8104
Criminalistics II
Professor Bensley
FRSC 8104 – DNA Objectives
1. Understand the basic principles of DNA
structure and analysis.
2. Recall the history and major steps in the
development of Forensic DNA Analyses
3. Describe and detail the methods of DNA
Extraction and Quantitation from an
evidentiary sample.
4. Describe and detail the steps involved in
the Polymerase Chain Reaction.
FRSC 8104 – DNA Objectives
5. Understand the importance of Short
Tandem Repeats in Forensic DNA Analysis.
6. Understand the function of the CODIS
system
7. Describe various techniques used in the
visualization of amplified DNA.
8. Convey the important concepts of
advanced DNA techniques to include YSTR and Mitochondrial DNA analyses.
FRSC 8104 – DNA Outline
I. Introduction and Overview
A. Sir Alec Jeffreys (1985)
1.
Colin Pitchfork
B. Results reported
in DNA analysis
FRSC 8104 – DNA Outline
II. Biological and Genetic Basics –
A. Basic DNA
Principles
Nucleotide Structure (dATP)
FRSC 8104 – DNA Outline
B. Chromosomes, Genes, DNA Markers
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.
FRSC 8104 – DNA Outline
C. DNA Marker Nomenclature
D. Population Variation
SEQUENCE POLYMORPHISM
LENGTH POLYMORPHISM
-------(AATG) (AATG) (AATG)-------3 repeats
------------(AATG) (AATG)-----------2 repeats
FRSC 8104 – DNA Outline
III. Historical Methods
A. Pre 1985
B. RFLP-Based DNA Typing
FRSC 8104 – DNA Outline
C. Early Based PCR Testing
1.
HLA DQ Alpha / DQA1
FRSC 8104 – DNA Outline
2.
Polymarker (PM+DQA1)
FRSC 8104 – DNA Outline
3.
D1S80
FRSC 8104 – DNA Outline
4.
Short Tandem Repeats
FRSC 8104 – DNA Outline
D. Silver vs. Fluorescent Staining Techniques
FRSC 8104 – DNA Outline
IV. DNA Extraction
A. Organic Extraction
B. Chelex Extraction
C. FTA Paper
D. Solid Phase Extraction
E. Differential Extraction
FRSC 8104 – DNA Outline
V. DNA Quantitation
A. Why?
B. How?
TAQMan
FRSC 8104 – DNA Outline
VI. DNA Amplification (PCR)
A.
B.
C.
D.
Introduction
Why PCR?
Issues with PCR
PCR Components
FRSC 8104 – DNA Outline
E. The Process of Replication
1.
2.
DNA Polymerase
Okazaki Fragments
http://rst.gsfc.nasa.gov/Sect20/DNA-20replication.gif
FRSC 8104 – DNA Outline
F.
G.
H.
I.
J.
Denaturation
Annealing
Extension
Multiplex PCR
PCR Inhibition
FRSC 8104 – DNA Outline
VII.Short Tandem Repeat Markers (STRs)
A. Repetitious DNA
B. Microsatellites or STR
FRSC 8104 – DNA Outline
C. 13 CODIS STR Loci
The Thirteen CODIS STRs and Their Probability of Identities
STR
African-American
Caucasian
D3S1358
0.094
0.075
vWA
0.063
0.062
FGA
0.033
0.036
THO1
0.109
0.081
TPOX
0.090
0.195
CSF1PO
0.081
0.112
D5S818
0.112
0.158
D13S317
0.136
0.085
D7S820
0.080
0.065
D8S1179
0.082
0.067
D21S11
0.034
0.039
D18S51
0.029
0.028
D16S539
0.070
0.089
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.
FRSC 8104 – DNA Outline
VIII.DNA Separation and Detection
A. Electrophoresis
1.
2.
Slab Gels
Capillary
B. DNA Detection
FRSC 8104 – DNA Outline
IX. Advanced Techniques
A. Y-STR
FRSC 8104 – DNA Outline
B. Mitochondrial DNA