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Optimization of InnoXtract™ extraction and purification system for DNA extraction from skeletal samples

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Abstract

The InnoXtract™ extraction and purification system is a purification method designed for DNA extraction from low-template samples, specifically rootless hair shafts. Its ability to successfully capture highly fragmented DNA suggests its suitability for use with other challenging sample types, including skeletal remains. However, the lysis and digestion parameters required modifications to successfully optimize the method for this sample type. A two-part digestion was developed utilizing a homebrew digestion buffer (0.5 M EDTA, 0.05% Tween 20, and 100 mM NaCl) and a supplemental lysis with the Hair Digestion Buffer included in the InnoXtract™ kit. Additionally, the magnetic bead volume was modified to improve DNA recovery from these challenging samples. With the altered protocol, the quality and quantity of DNA recovered from InnoXtract™ extracts were comparable to another commercial skeletal extraction method (PrepFiler™ BTA). This modified extraction method successfully purified sufficient amounts of quality DNA from a variety of skeletal samples to produce complete STR profiles. Successful STR typing from surface decomposition, burned, cremated, buried, and embalmed remains indicates the potential of this new method for challenging human identification and missing-person cases.

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Acknowledgements

The authors would like to thank the Southeast Texas Applied Forensic Science Facility (a willed body donor program at Sam Houston State University), the donors, and their loved ones, without whom this research would not be possible. The authors would also like to thank InnoGenomics Technologies for providing valuable feedback and technical support.

Funding

This project was partially funded by Award # 1739805 through the Center for Advanced Research in Forensic Science (CARFS), an NSF-funded IUCRC at Florida International University. The opinions, findings, conclusions, or recommendations expressed in this article are those of the authors.

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Authors and Affiliations

  1. Department of Forensic Science, College of Criminal Justice, Sam Houston State University, 1003 Bowers Blvd., TX, 77341, Huntsville, USA

    Jennifer Snedeker, Sheree Hughes & Rachel Houston

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  1. Jennifer Snedeker
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  2. Sheree Hughes
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  3. Rachel Houston
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Corresponding author

Correspondence to Jennifer Snedeker.

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Ethical approval

For the skeletal samples used in this study, Code 45 of the US Federal Regulations part 46102(f) exempts the requirement for Institutional Review Board (IRB) approval regarding the use of human cadaveric samples. All procedures were in accordance with the 1964 Helsinki Declaration and its later amendments.

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The authors declare no competing interests.

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Snedeker, J., Hughes, S. & Houston, R. Optimization of InnoXtract™ extraction and purification system for DNA extraction from skeletal samples. Int J Legal Med 137, 949–959 (2023). https://doi.org/10.1007/s00414-023-02980-9

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  • DOI: https://doi.org/10.1007/s00414-023-02980-9

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