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. 2021 Jan 8:3:100131.
doi: 10.1016/j.fsisyn.2020.100131. eCollection 2021.

Improved autosomal STR typing of degraded femur samples extracted using a custom demineralization buffer and DNA IQ™

Gayvelline C Calacal  1 , Bea G Gallardo  1 , Dame Loveliness T Apaga  1 , Maria Corazon A De Ungria  1
Affiliations

Improved autosomal STR typing of degraded femur samples extracted using a custom demineralization buffer and DNA IQ™

Gayvelline C Calacal et al. Forensic Sci Int Synerg. .

Abstract

Bone samples are utilized as a source of DNA in disaster victim identification and human remains investigations. However, DNA recovery from bones is time consuming and prone to contamination. A logical approach for postmortem identification is to validate efficient DNA extraction methods requiring less bone material using improved molecular kits with less hands-on time and workflows that facilitate faster turn-around time for processing. In this study, we evaluated DNA yield and amplification efficiency of DNA extracts using a new custom bone demineralization buffer (DMB; Promega) and extracted via manual and automated DNA IQ™ workflows. Including the demineralization step, the bone protocol can be completed in ∼4 h and even less with minimal sample handling when automated. Overall, a rapid and simple DNA extraction with improved allele recovery was validated using degraded bone samples exposed to tropical environments and post-disaster recovery as well as adverse conditions of embalming prior to internment.

Keywords: Automation; Autosomal STR; Bone; DNA extraction; Demineralization; Forensic.

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Conflict of interest statement

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Figures

Fig. 1
Fig. 1
A. DNA yield and B. amplification efficiency per DNA extraction method (manual workflow) using 16 femur samples. Mean DNA yield computed for duplicate extracts except for femur samples that were obtained from a Disaster Victim Identification (DVI) investigation. Horizontal lines indicate mean allele recovery for each DNA extraction protocol.
Fig. 2
Fig. 2
A. DNA yield and B. amplification efficiency per DNA extraction method (automated workflow) using 16 femur samples. Mean DNA yield computed for duplicate extracts except for femur samples that were obtained from a Disaster Victim Identification (DVI) investigation. Horizontal lines indicate mean allele recovery for each DNA extraction protocol.
Fig. 3
Fig. 3
A. DNA concentration B. DNA yield and C. amplification efficiency per initial bone powder weight extracted with DMB/DNA IQ™ (manual workflow) using 16 femur samples. Mean DNA yield computed for duplicate extracts except for femur samples that were obtained from a Disaster Victim Identification (DVI) investigation. Horizontal lines indicate mean allele recovery for each bone weight (mg).
Fig. 4
Fig. 4
A. DNA yield and B. amplification efficiency after pre-treatment with a custom demineralization buffer (DMB) then extracted using a manual DNA IQ™ or an automated Maxwell® 16 LEV workflow using 16 femur samples. Mean DNA yield computed for duplicate extracts except for femur samples that were obtained from a Disaster Victim Identification (DVI) investigation. Horizontal lines indicate mean allele recovery for each DNA extraction protocol.
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References

    1. Ward J. Best practice recommendations for the establishment of a national DNA identification program for missing persons: a global perspective. Forensic Sci. Int.: Genetics Supplement Series. 2017;6:e43–e45.
    1. Watherston J., McNevin D., Gahan M.E., Bruce D., Ward J. Current and emerging tools for the recovery of genetic information from postmortem samples: new directions for disaster victim identification. Forensic Sci. Int.: Genetics. 2018;37:270–282. - PubMed
    1. Edson S.M., McMahon T.P. Extraction of DNA from skeletal remains. In: Goodwin W., editor. Forensic DNA Typing Protocols. Humana Press; New York, NY: 2016. pp. 69–87.
    1. Loreille O.M., Diegoli T.M., Irwin J.A., Coble M.D., Parsons T.J. High efficiency DNA extraction from bone by total demineralization. Forensic Sci. Int.: Genetics. 2007;1(2):191–195. - PubMed
    1. Lee H.Y., Park M.J., Kim N.Y., Sim J.E., Yang W.I., Shin K.J. Simple and highly effective DNA extraction methods from old skeletal remains using silica columns. Forensic Sci. Int.: Genetics. 2010;4(5):275–280. - PubMed

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