Unidentified human remains are often associated with tragic events, such as fires, terrorist attacks, natural disasters, war conflicts, genocide, airline crashes, homicide, and human rights violations under oppressive totalitarian regimes. In these situations, extensive damage to soft tissues often precludes the use of such biological samples in the identification process. In contrast, bone material is the most resilient, viable sample type for DNA testing. DNA recovered from bone often is degraded and in low quantities due to the effects of human decomposition, environmental exposure, and the passage of time. The complexities of bone microstructure and its rigid nature make skeletal remains one of the most challenging sample types for DNA testing.
Table of Contents
1. Missing persons and unidentified human remains: The world's silent mass disaster 2. Challenges in forensic genetic investigations of decomposed or skeletonized human remains: Environmental exposure, DNA degradation, inhibitors, and low copy number (LCN)
I. Guidelines and best practices for handling and processing human skeletal remains for genetic studies 3. Facilities design and workflow considerations for processing unidentified human skeletal remains 4. Location, recovery, and excavation of human remains for forensic testing 5. Skeletal microstructure, bone diagenesis, optimal sample selection, and pre-processing preparation techniques for DNA testing 6. DNA extraction methods for human skeletal remains 7. Quantitative and qualitative assessment of DNA recovered from human skeletal remains
II. Types of DNA markers and applications for identification 8. Autosomal short tandem repeat (STR) profiling of human 9. Y-chromosome analysis for unidentified human remains (UHR) investigations 10. Mitochondrial DNA and its use in the forensic analysis of skeletal material 11. X-chromosome short tandem repeats (X-STRs): Applications for human remains identification 12. Single nucleotide polymorphisms (SNPs): Ancestry-, phenotype-, and identity-informative SNPs 13. Diallelic Markers: INDELs and INNULs
III. Traditional platforms, alternative strategies, and emerging technologies for DNA analysis of human skeletal remains 14. Genotyping and sequencing of DNA recovered from human skeletal remains using capillary electrophoresis (CE) 15. Rapid DNA identification of human skeletal remains 16. Emerging technologies for DNA analysis of challenged samples
IV. Analysis of genetic data recovered from skeletonized human remains 17. Best practices in the development and effective use of a forensic DNA database for identification of missing persons and unidentified human remains 18. Software and database functionality for direct identification and kinship analysis: The Mass Fatality Identification System (M-FISys) 19. Bioinformatic tools for interrogating DNA recovered from human skeletal remains 20. The emerging discipline of forensic genetic genealogy
V. Complementary and multidisciplinary approaches to assist in identification of unidentified human skeletal remains 21. Forensic anthropology in a DNA world: How anthropological methods complement DNA-based identification of human remains 22. Generation of a personal chemical profile from skeletonized human remains 23. Forensic odontology: Historical perspectives and current applications for identification of human remains 24. Forensic facial reconstruction of skeletonized and highly decomposed human remains
Authors
Angie Ambers Director of Forensic Genetics at Institute for Human Identification, LMU College of Dental Medicine, Tennessee, USA. Dr. Angie Ambers is an internationally recognized DNA expert and Assistant Director of the Henry C. Lee Institute of Forensic Science, a world-class institute that specializes in interdisciplinary research, training, testing, casework consulting, and education in forensic science. Dr. Ambers also holds an Associate Professor (Forensic DNA) appointment in the Henry C. Lee College of Criminal Justice and Forensic Sciences at the University of New Haven, teaching forensic biology and DNA analysis methods, in addition to specialty courses on advanced DNA topics. She has a Ph.D. in molecular biology (with emphasis in forensic genetics and human identification) as well as master's degrees both in forensic genetics and in criminology. She worked as a forensic geneticist for the University of North Texas (UNT) Center for Human Identification for 8 years and, prior to pursing her doctorate, was lead DNA analyst and lab manager of UNT's DNA Sequencing Core Facility. Her doctoral research involved an investigation of methods (e.g., whole genome amplification, DNA repair) for improving autosomal and Y-STR typing of degraded and low copy number (LCN) DNA from human skeletal remains and environmentally damaged biological materials. Her master's thesis research included development and optimization of a DNA-based multiplex screening tool for genetics-based separation of fragmented and commingled skeletal remains in mass graves.Dr. Ambers specializes in characterization and identification of contemporary, historical, and archaeological human skeletal remains. Her casework has involved DNA testing of an American Civil War guerrilla scout; several Finnish World War II soldiers; unidentified late-19th century skeletal remains discovered by a construction crew in Deadwood, South Dakota; unidentified skeletal remains of Special Operations soldiers killed during the 1974 Turkish invasion of Cyprus; skeletal remains exhumed from Prague Castle in the Czech Republic; skeletal remains of soldiers from the Seven Years' War (1756-1763); bone samples purported to belong to a member of Jesse James gang (killed during a bank robbery in 1876); and the exhumed remains of the wife of a Yale medical school professor. She co-presented a workshop on "Advanced Methods for DNA-based Analysis of Skeletal Remains� at the 26th International Symposium on Human Identification (ISHI), and has been an invited speaker at an international bone workshop/conference in Prague, Czech Republic. In 2017, she traveled twice to India to train scientists from various Indian states and the Maldives Police Service on the processing of bone samples in forensic DNA casework. During her visit to India, she performed autosomal STR analysis on human skeletal remains discovered along a hiking route in the Himalayas (in the northern state of Himachal Pradesh) to assist local officials in the investigation of a missing persons case. Additionally, she performed DNA analysis on a female homicide victim recovered from a clandestine grave in New Delhi.
Dr. Ambers' casework and research has been published in various peer-reviewed journals, including Forensic Science International: Genetics (FSI:Genetics), Forensic Science International, International Journal of Legal Medicine, Legal Medicine, BMC Genomics, the Croatian Medical Journal, The Journal of Heredity, and Journal of Biological and Clinical Anthropology (Anthropologischer Anzeiger). Her work has received press in numerous local and national newspapers (including The Washington Times, NBC News, Criminal Legal News, The Root) and has been featured on several podcasts (Truth and Justice, Crime Waves).
Among her most recent casework (a collaboration with the Texas Historical Commission) includes DNA analyses of human skeletal remains associated with the French explorer La Salle's last expedition. This case involved massively parallel sequencing (MPS) of DNA from two adult male skeletons recovered from the La Belle shipwreck, skeletal remains recovered from a mass grave at Fort St. Louis, and the putative remains of Sieur de Marle. Additionally, Dr. Ambers performed DNA extractions and mitochondrial DNA (mtDNA) sequencing on numerous sets of skeletal remains associated with Spanish royalty and the House of Aragon (recovered from tombs within the Royal Pantheon of San Juan de la Pe�a archaeological site in Spain). She is currently working on the putative remains of William Townsend Washington, the nephew of former U.S. President George Washington.
In addition to skeletal remains cases and research, Dr. Ambers collaborated with the National Institute of Justice (NIJ) and the Forensic Technology Center of Excellence (FTCoE) to develop and disseminate a formal report on the use of Familial DNA Searching (FDS) in casework. She is an active cold case consultant, an advocate of post-conviction DNA testing, and an educator/advisor on DNA testing or re-testing of old, degraded, or challenging evidentiary samples. Dr. Ambers also was the 2017-2018 Project Lead on a U.S. State Department grant to combat human trafficking in Central America through the application of forensics. As part of the program objectives, she traveled to three Northern Triangle countries (Guatemala, El Salvador, Honduras) to perform gap assessments of government laboratories and train personnel in forensic DNA analysis, with the goal of promoting quality casework methods based on ISO 17025 standards. As Project Lead, she helped train forensic DNA scientists at the Instituto Nacional de Ciencias Forenses de Guatemala (INACIF, the National Institute of Forensic Sciences of Guatemala) in Guatemala City, Guatemala; the Instituto de Medicina Legal (Institute of Legal Medicine) in San Salvador, El Salvador; and the Instituto de Medicina Forenses: Centro de Medicina Legal y Ciencias Forenses (the Institute of Forensic Medicine: Center of Legal Medicine and Forensic Sciences) in Tegucigalpa, Honduras. In addition to providing both lecture and
