Abstract
STR analysis of DNA extracted from skeletal samples can play an important role in the identification of missing persons. Here we present a method for the extraction of DNA from skeletal samples involving complete demineralization and digestion of the sample, followed by purification by silica binding. This method, together with the multiplex STR typing approach also presented, has proven highly successful in the recovery of DNA profiles from degraded, aged skeletal remains from a wide range of environmental contexts. The methodological steps presented include bone decontamination and grinding, DNA extraction, repurification in the case of highly inhibited samples, quantification, STR multiplex amplification, and profile reporting guidelines. However, the conditions applied for amplification and the criteria for allele calling and profile submission must be based on the results of each laboratory’s internal validation experiments involving the type of samples relevant to the project at hand. The methods presented here have permitted large-scale DNA-based identification of persons missing from mass disasters and armed conflict.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Alonso A, Martin P, Albarran C et al (2005) Challenges of DNA profiling in mass disaster investigations. Croat Med J 46:540–548.
National Research Council of the National Academies (2009) Strengthening forensic science in the United States: a path forward. National Academies Press, Washington DC.
Huffine E, Crews J, Kennedy B et al (2001) Mass identification of persons missing from the break-up of the former Yugoslavia: structure, function, and role of the International Commission on Missing Persons. Croat Med J 42:271–275.
Rohland N and Hofreiter M (2007) Comparison and optimization of ancient DNA extraction. Biotechniques 42:343–352.
Holland MM, Fisher DL, Mitchell LG (1993) Mitochondrial DNA sequence analysis of human skeletal remains: identification of remains from the Vietnam War. J Forensic Sci 38:542–553.
Fisher DL, Holland MM, Mitchell L (1993) Extraction, evaluation, and amplification of DNA from decalcified and undecalcified United States Civil War bone. J Forensic Sci 38:60–68.
Davoren J, Vanek D, Konjhodzić R et al (2007) Highly effective DNA extraction method for nuclear short tandem repeat testing of skeletal remains from mass graves. Croat Med J 48:478–485.
Parsons TJ, Huel R, Davoren J et al (2007) Application of novel ‘mini-amplicon’ STR multiplexes to high volume casework on degraded skeletal remains. FSI:Genetics 1:175–179.
Miloš A, Selmanović A, Smajlović et al (2007) Success rates of nuclear short tandem repeat typing from different skeletal elements. Croat Med J 48:486–493.
Mundorff AZ, Bartelink EJ and Mar-Cash E (2009) DNA preservation in skeletal elements from the World Trade Center disaster: recommendations for mass fatality management. J Forensic Sci 54:739–745.
Edson SM, Ross JP, Michael D. Coble MD et al (2004) Naming the dead: confronting the realities of the rapid identification of degraded skeletal remains. Forensic Sci Rev 16:63–90.
Salamon M, Tuross N, Arensburg B et al (2005) Relatively well preserved DNA is present in the crystal aggregates of fossil bones. Proc Natl Acad Sci 102:13783–13788.
Loreille OM, Diegoli TM, Irwin JA et al (2007) High efficiency extraction of DNA from bone by total demineralization. Forensic Sci Int Genet 1: 191–195.
Yang DY, Eng B, Waye JS et al (1998) Technical note: improved DNA extraction from ancient bones using silica-based spin columns. Am J Phys Anthropol 105:539–543.
Lee HY, Park MJ, Kim NY et al (2010) Simple and highly effective DNA extraction methods from old skeletal remains using silica columns. Forensic Sci Int Genet 4:275–280.
Seo SB, Zhang A, Kim HY (2010) Technical note: efficiency of total demineralization and ion-exchange column for DNA extraction from bone. Am J Phys Anthropol 141:158–162.
Acknowledgments
Many ICMP staff members have been involved in the development, validation, and/or long-term use of the protocols presented here; their excellent work and dedication is much appreciated. We particularly acknowledge the managerial and other assistance of Adnan Rizvić, Adam Boys, and Kathryne Bomberger.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Huel, R., Amory, S., Bilić, A., Vidović, S., Jasaragić, E., Parsons, T.J. (2012). DNA Extraction from Aged Skeletal Samples for STR Typing by Capillary Electrophoresis. In: Alonso, A. (eds) DNA Electrophoresis Protocols for Forensic Genetics. Methods in Molecular Biology, vol 830. Humana Press. https://doi.org/10.1007/978-1-61779-461-2_13
Download citation
DOI: https://doi.org/10.1007/978-1-61779-461-2_13
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-460-5
Online ISBN: 978-1-61779-461-2
eBook Packages: Springer Protocols