PCR Machine

Multiplex PCR Sensitivities to levels less than 1 ng of DNA Over 10 Markers Can Be Copied at Once Ability to Handle Mixtures and Degraded Samples Different Fluorescent Dyes Used to Distinguish STR Alleles with Overlapping Size Ranges

How PCR works

Capillary electrophoresis with multi-color detection capabilities ABI PRISM® 310 Genetic Analyzer Automated gel pouring Automated sample injection Capillary electrophoresis with multi-color detection capabilities

A case of DNA Match Inclusion Suspected Control

DNA Profiling CAN’T ESCAPE DNA TEST

DNA Examinations In the Absence of Mother or Father’s Biological Sample Mt-DNA Analysis Y-Chromosome DNA Analysis

Human Genome 23 Pairs of Chromosomes + mtDNA Located in cell nucleus http://www.ncbi.nlm.nih.gov/genome/guide/ 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 X Y Autosomes 2 copies per cell Located in mitochondria (multiple copies in cell cytoplasm) mtDNA 16,569 bp Mitochondrial DNA Figure 2.3 The human genome contained in every cell consists of 23 pairs of chromosomes and a small circular genome known as mitochondrial DNA. Chromosomes 1-22 are numbered according to their relative size and occur in single copy pairs within a cell’s nucleus with one copy being inherited from one’s mother and the other copy coming from one’s father. Sex-chromosomes are either X,Y for males or X,X for females. Mitochondrial DNA is inherited only from one’s mother and is located in the mitochondria with hundreds of copies per cell. Together the nuclear DNA material amounts to over 3 billion base pairs (bp) while mitochondrial DNA is only about 16,569 bp in length. Sex-chromosomes Nuclear DNA 3.2 billion bp 100s of copies per cell

*Minimal haplotype loci names are hyperlinked to STR fact sheets  

(passed on complete, but only by sons) Lineage Markers Autosomal (passed on in part, from all ancestors) Figure 9.1 Illustration of inheritance patterns from recombining autosomal genetic markers and the lineage markers from the Y-chromosome and mitochondrial DNA. Y-Chromosome (passed on complete, but only by sons) Mitochondrial (passed on complete, but only by daughters)

Tracing paternal lineage

Y Chromosome STR Marker Study For Tracing Paternal Lineages Forensically Important Markers: DYS389 (I and II), DYS390, DYS19, DYS385, DYS391, DYS392

Modern Use of Y-STR Testing Captured December 13, 2003 Uday and Qusay Hussein Killed July 22, 2003 Matching Y-STR Haplotype Used to Confirm Identity (along with allele sharing from autosomal STRs) Is this man really Sadaam Hussein?

A CASE STUDY FATHER Son-I Son-II Son-III MISSING

LADDER QUESTIONED EXHIBIT FATHER Electropherogram SON-I SON-II

NATIONAL DNA ANALYSIS CENTER CENTRAL FORENSIC SCIENCE LABORATORY Ministry of Home Affairs, Govt. of India 30 Gorachand Road, Park Circus, Kolkata- 700014, West Bengal FORENSIC DNA EXAMINATION REPORT Report No.: CFSL(K)/EE/2005(M.P.)-XXX Date: XXX 2007

Mitochondrial DNA (mtDNA)

Mitochondrial DNA (mtDNA) Cons Single-cell sensitivity because each cell contains ~1000 mitochondria = very high contamination risk! Heteroplasmy - more than one mtDNA type manifesting in different tissues in the same individual Lower power of discrimination - maternal relatives all share the same mtDNA

Dynamics of mt-DNA inheritance Typing a SNP with Microchip

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After a decade or so… Here is my sequence