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The Effects of Natural Mummification on the Recovery and Degradation of DNA The University of Southern Mississippi The Aquila Digital Community Honors Theses Honors College 5-2021 Unraveling the Mummy: The Effects of Natural Mummification on the Recovery and Degradation of DNA Marissa Gandolfo-Muller Follow this and additional works at: https://aquila.usm.edu/honors_theses Part of the Forensic Science and Technology Commons Recommended Citation Gandolfo-Muller, Marissa, "Unraveling the Mummy: The Effects of Natural Mummification on the Recovery and Degradation of DNA" (2021). Honors Theses. 779. https://aquila.usm.edu/honors_theses/779 This Honors College Thesis is brought to you for free and open access by the Honors College at The Aquila Digital Community. It has been accepted for inclusion in Honors Theses by an authorized administrator of The Aquila Digital Community. For more information, please contact [email protected]. Unraveling the Mummy: The Effects of Natural Mummification on the Recovery and Degradation of DNA by Marissa Gandolfo-Muller A Thesis Submitted to the Honors College of The University of Southern Mississippi in Partial Fulfillment of Honors Requirements May 2021 iii Approved by: Dr. Marie Danforth, Ph.D., Thesis Advisor, School of Social Science and Global Studies Dr. Kuppareddi Balamurugan, Ph.D., Thesis Co- Advisor, School of Criminal Justice, Forensic Science and Security Dr. Bridget Hayden, Ph.D., Interim Director, School of Criminal Justice, Forensic Science and Security Ellen Weinauer, Ph.D., Dean Honors College iii ABSTRACT As demonstrated through casework and research studies (Hawass et al., 2010; Gielda & Rigg, 2017), anthropogenic mummification and modern-day embalming can expedite degradation of DNA. Current research in the field of forensic mummification is sparse and little research has been done on quantifying naturally mummified DNA (Leccia et al., 2018; Shved et al., 2014). This research focuses on observing and quantifying the differences in the recovery and degradation of DNA from specimens that have been naturally mummified. This research on natural, forensic mummies is a blend of experimental archeology and postmortem DNA analysis. In this study, two control specimens and seven experimental specimens were used. Of the nine specimens, three of the specimens partially mummified, three specimens showed signs of superficial mummification and three specimens naturally decomposed. The specimens exposed to salt of neutral pH and cold temperatures, well known preservations of tissue and DNA, had greater DNA yield and lower rates of postmortem DNA degradation. The specimens exposed to UV radiation, alkaline pHs, and high temperatures showed lower DNA yield and higher levels of DNA degradation. The results of this research could make contributions to the fields of forensic identification and forensic anthropology, specifically, cold cases, victim identification in mass disasters and wars, and identification of genetic abnormalities within large gravesites through DNA analysis. Keywords: natural mummification, experimental archeology, DNA analysis, DNA recovery, DNA degradation iv DEDICATION To my fellow scholars with unwavering passions for Egyptology, archaeology, and forensic science: death is only the beginning. v ACKNOWLEDGMENTS First and foremost, thank you to Dr. Danforth for her guidance and endless positivity throughout this process. I am forever thankful for your mentorship and encouragement in all my academic endeavors. I hope that whenever you think of chickens, you will also think of me. Thank you to Dr. Balamurugan for the continuous use of his laboratory and knowledge of DNA. Thank you to the Honors College for unwavering patience and encouragement during the writing process and for believing in me through my journey as a Foundations student to a Keystone scholar. Thank you for also financially supporting the advancement of my thesis research. Thank you to the Drapeau Center for Undergraduate Research for also funding my research through the Eagle SPUR Program. Thank you to my mom and dad for fostering my love of Ancient Egypt. Thank you to my aunt for Starbucks gifts cards that got me through the long writing sessions. Thank you to everyone who has patiently listened to me babble about mummified chickens over the last year. Thank you to Julia, Maggie, and Helen for providing coffee, friendship, and a quiet space to be thesis writing buddies at. A special thank you to Anthony and Chandler for being my chicken couriers. vi TABLE OF CONTENTS LIST OF TABLES ............................................................................................................. ix LIST OF ILLUSTRATIONS .............................................................................................. x LIST OF ABBREVIATIONS ............................................................................................ xi INTRODUCTION ........................................................................................ 1 LITERATURE REVIEW ........................................................................... 3 Mummification in Cultural Settings ............................................................................... 3 The Process of Mummification ....................................................................................... 7 The Basics of DNA ....................................................................................................... 10 Experimentation with Mummies and DNA .................................................................. 13 METHODS AND MATERIALS ............................................................ 15 Recreated Natural Mummification Settings.................................................................. 15 Open Air Natural Decomposition Specimen ............................................................ 15 Soil Natural Decomposition Specimen ..................................................................... 16 Desert Natural Mummification Specimen ................................................................ 16 Air-based Natural Mummification Specimen ........................................................... 16 Rock Salt Natural Mummification Specimen ........................................................... 17 Saline Lake Natural Mummification Specimen ........................................................ 17 Frozen Natural Mummification Specimen................................................................ 17 Cave-based Natural Mummification Specimen ........................................................ 18 viii Bog Body Natural Mummification Specimen .......................................................... 18 Sample Collection ..................................................................................................... 19 DNA Analysis ............................................................................................................... 19 DNA Reagents .......................................................................................................... 19 Organic DNA Extraction .......................................................................................... 21 Gel Electrophoresis ................................................................................................... 22 Nanodrop Spectrophotometer ................................................................................... 22 Second TE Washing .................................................................................................. 23 RNase Treatment ...................................................................................................... 23 Ethanol Precipitation of DNA ................................................................................... 23 RESULTS ................................................................................................ 25 Mummification Results ................................................................................................. 25 DNA Analysis Results .................................................................................................. 27 DISCUSSION ........................................................................................... 34 CONCLUSION ....................................................................................... 36 Nanodrop Reading After Organic Extraction ......................................... 39 Nanodrop Reading After RNase Treatment ............................................ 40 Nanodrop Reading After Ethanol Precipitation ...................................... 41 REFERENCES ................................................................................................................. 42 viii LIST OF TABLES Table 1: Degrees of mummification ................................................................................. 25 Table 2: Specimen mummification results ....................................................................... 26 Table 3: Specimen weight changes ................................................................................... 26 Table 4: Sample key.......................................................................................................... 27 Table 5: Final DNA yield calculations ............................................................................. 31 ix LIST OF ILLUSTRATIONS Figure 1: DNA Structure ................................................................................................... 11 Figure 2: Gel 1 showing the DNA quantity and quality ................................................... 28 Figure 3:
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