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ELEquant: a developmental framework and validation of forensic and conservation real-time PCR assays

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Abstract

A framework for the development and validation of a qPCR assay for species identification and DNA quantification for conservation and forensic purposes is presented. Elephants are commonly poached for their ivory tusks, which is the primary driving force behind their endangered status. In addition to poaching and trade, habitat loss due to logging and mining has also resulted in loss of elephants. Crimes against animals can be deterred and/or further prosecution sought through testing with forensic genetic techniques. The creation of novel genetic assays can greatly impact wildlife forensic science investigations in identifying the species. Molecular genetic techniques can help enforce conservation efforts; however, they must be properly developed and validated to be of evidentiary quality for court systems. African and Asian elephant buccal cells were used as model in this work. The assay provides a method to differentiate biological fluids of both genera of elephants simultaneously. It can be used for identification of elephant derived products and presents valuable quantification for optimized further testing, such as microsatellite detection.

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Acknowledgements

We thank Merideth Fayman, Caitlin Hoey, and Meredith Rohrbaugh for early contributions to this research project. This work would not be possible without DNA sources from Six Flags: Safari Off Road Adventure, the Cincinnati Zoo, the St. Louis Zoo, the Northeast Wildlife DNA Laboratory, Quakertown Veterinary Clinic, and the Brandywine Zoo. Thank you for your contributions. Thank you to Arcadia University’s Master of Forensic Science Program and the Department of Biological and Physical Sciences at Keystone College for providing funds towards this project.

Funding

This work has been supported by Arcadia University’s Master of Forensic Science program, Keystone College’s Department of Biological and Physical Sciences, and University of the Sciences’ Department of Biological Sciences.

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Authors and Affiliations

  1. Department of Biological and Physical Sciences, Keystone College, One College Green, La Plume, PA, 18440, USA

    Jillian Conte

  2. Department of Biological Sciences, University of the Sciences, Philadelphia, PA, USA

    Jillian Conte

  3. Department of Chemistry, Physics and Forensic Science, Arcadia University, 450 S. Easton Road, Glenside, PA, 19038, USA

    Margret J. Potoczniak & Shanan S. Tobe

  4. Armed Forces Medical Examiner System’s Armed Forces DNA Identification Laboratory (AFMES-AFDIL), Dover Air Force Base, Dover, DE, USA

    Courtney Mower

  5. ARP Sciences, LLC, Rockville, MD, USA

    Courtney Mower

  6. College of Science and Engineering, Flinders University, Adelaide, SA, 5042, Australia

    Shanan S. Tobe

Authors
  1. Jillian Conte
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  2. Margret J. Potoczniak
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  3. Courtney Mower
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  4. Shanan S. Tobe
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Correspondence to Jillian Conte.

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All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

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Conte, J., Potoczniak, M.J., Mower, C. et al. ELEquant: a developmental framework and validation of forensic and conservation real-time PCR assays. Mol Biol Rep 46, 2093–2100 (2019). https://doi.org/10.1007/s11033-019-04660-7

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  • DOI: https://doi.org/10.1007/s11033-019-04660-7

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