y str markers

y str markers

1. Ministry of Higher education & scientific Research Al- Mustaqbal University Collage   STR markers of Y chromosome By Dr. Mohammed Zuhair Al- Murib Ph.D in Human Genetic & Forensic Medicin Department of Medical lab. Techniques  

2. Presentation Outline Structure of the Y Chromosome Advantages of Y Chromosome Markers Commonly Used of Y Chromosome Markers Y STR Multiplex Assays Applications of Y STR in forensic medicine

3. Structure of the Y Chromosome p q

4. Why the Y Chromosome Markers ? forensic investigations genealogical purposes evolutionary studies Advantages to human identity testing paternal lineages

5. Autosomal (passed on in part, from all ancestors) Y-Chromosome (passed on complete, but only by sons) Mitochondrial (passed on complete, but only by daughters) Lineage Markers Figure 9.1, J.M. Butler (2005) Forensic DNA Typing , 2 nd Edition © 2005 Elsevier Academic Press Vertical heredity

6. Y Chromosome DNA Markers Microsatellites is a tract of repetitive DNA in which certain DNA motifs (ranging in length from two to five base pairs ) are repeated, typically 5–50 times. Microsatellites occur at thousands of locations within an organism's genome. STRs mostly tetranucleotide repeats Bi-allelic markers (unique event polymorphisms--UEP) SNPs ( single nucleotide polymorphisms ) Y A lu p olymorphism (YAP) or other insertions/deletions (“ indels ”) Minisatellite is a tract of repetitive DNA in which certain DNA motifs (ranging in length from 10–60 base pairs ) are typically repeated 5-50 times.

7. Y Chromosome DNA Markers short tandem repeat (STR) in DNA occurs when: a pattern of two or more nucleotides are repeated the repeated sequences are directly adjacent to each other. The pattern can range in length from 2 to 5 base pairs (bp) is typically located in the non-coding intron region. A short tandem repeat polymorphism (STRP) occurs when homologous STR loci differ in the number of repeats between individuals.

8. THE PROJECT OF ERA

9. Y-Chromosome Consortium

10. N ational C enter for B iotechnology I nformation Search database NCBI

11. THE PROJECT OF ERA

12. STR DataBase

13. Primer Sequences From Y STR Database http://ystr.charite.de/

14. STR loci

15. Y Chromosomal Adam

16. Applications of Y STR in forensic medicine Sources of Biological Evidence Blood Semen Saliva Urine Hair Teeth Bone Tissue Blood stain Only a very small amount of blood is needed to obtain a DNA profile

17. Sample Obtained from Crime Scene or Paternity Investigation DNA Extraction DNA Quantitation PCR Amplification of Multiple STR markers Biology Separation and Detection of PCR Products (STR Alleles) Technology Sample Genotype Determination Genetics Comparison of Sample Genotype to Other Sample Results If match occurs, comparison of DNA profile to population databases Generation of Case Report with Probability of Random Match

18. Steps in DNA Analysis DNA Database STR Typing DNA Extraction Multiplex PCR Amplification Male: 13,14-15,16-12,13-10,13-15,16 Interpretation of Results Sample Collection & Storage Buccal swab Blood Stain DNA Quantitation Slot Blot 1 ng 0.3 ng 1 ng 1 ng 0.7 ng 0.5 ng 0.5 ng No DNA

19. Capillaries Electrodes for Injection Figure 14.4, J.M. Butler (2005) Forensic DNA Typing , 2 nd Edition © 2005 Elsevier Academic Press

20. Scanned Gel Image Capillary Electropherogram 8 repeats 9 repeats

21. AMEL D3S1358 TH01 TPOX D2S1338 D19S433 FGA D21S11 D18S51 CSF1PO D16S539 D7S820 D13S317 D5S818 VWA D8S1179 Information is tied together with multiplex PCR and data analysis AmpFlSTR ® Identifiler ™ (Applied Biosystems)

22. Family Inheritance of STR Alleles (D13S317) Father Child #1 Child #2 Child #3 Mother PCR product size (bp) 11 14 11 12 14 8 14 12 12 8 Adam Marshall Katy Father Mother PATERNITY TESTING

23. Previous studies Chiaroni 2009 Al-Zahery 2011 Tofanelli 2009

24. “We got him!” Saddam Hussein’s capture verified with DNA testing December 14, 2003 DNA Profile Source: www.cnn.com; The Scientist Dec 19, 2003

25. Modern Use of Y-STR Testing Captured December 13, 2003 Is this man really Sadaam Hussein? Uday and Qusay Hussein Killed July 22, 2003 Matching Y-STR Haplotype Used to Confirm Identity (along with allele sharing from autosomal STRs) Butler, J.M. (2005) Forensic DNA Typing, 2 nd Edition, Box 23.1, p. 534

26. Uday and Qusay Hussein Killed July 22, 2003 Captured December 13, 2003 Source: www.cnn.com Suspected Saddam Portion of DNA Profile “A Forensic Paternity Test” Y-chromosome tests were also used to verify male lineage… DNA Used as the Ultimate Biometric for Confirming Saddam’s Identity

27. Anastasia Romanova

28. Tomb of Unknown Soldier Armed Forces DNA Identification Laboratory (Rockville, MD) Identification of Skeletal Remains from Previous Military Conflicts Identification of Michael J. Blassie - Vietnam Tomb of the Unknown >2100 Unaccounted for from Southeast Asia >8000 Unaccounted for from Korea >75,000 Unaccounted for from WWII

29. 30 THANK YOU FOR ATTENTION