Date of Submission

5-2026

Document Type

Thesis

Degree Name

Master of Science in Forensic Science

Department

Criminal Justice

Advisor

Ashley G. Morgan, Ph.D.

Committee Member

Laura G. Combs, Ph.D.

Committee Member

Angelique P. Corthals, Ph.D.

Keywords

Submerged Skeletal Remains, DNA Degradation, Sus Scrofa Domesticus, Proteinase K, DTT (Modified Extraction Protocol), Post-Mortem Submersion Interval (PMSI)

MeSH

Bones, DNA, Swine, Cell fractionation, Real-time polymerase chain reaction, Forensic anthropology

LCSH

Skeletal remains, DNA, Swine, Enzymes, Proteins, Postmortem changes

Abstract

Recovering DNA from submerged skeletal remains for the positive identification of individuals is a challenging task. Factors like water temperature and depths as well as the presence of coverings on the victim can impact the rate of decomposition which in tum can impact the degree of DNA degradation. The currently available literature suffers from a lack of studies that examine how the quantity of DNA in submerged skeletal remains changes over time, but a few studies have indicated that the first week is the most successful for recovering DNA. The current study examined how DNA quantities changed over four weeks in juvenile Sus scrofa domesticus femurs submerged in fresh water. Multiple femurs were added to one tank with approximately one inch of sand substrate on the bottom of the tank. One femur was recovered from the water every 24 hours for one week, then weekly for one month. Ten samples were taken from each submerged femur with half of the samples undergoing the vendor-specified extraction protocol and the other half undergoing a modified protocol which substituted Proteinase K with an equivalent volume of dithiothreitol (DTT). The recovery between the protocols was compared to determine which protocol yielded a higher quantity of DNA. Samples were recovered through a combination of cutting and powdering. Genomic DNA was extracted from the powdered samples by solid-phase methods through the use of magnetic silica beads and assessed by real­ time PCR quantification. Quantifiable DNA was recovered for every time point through Day 28 of submersion. The largest decrease in DNA yields was observed within the first 3 days after the bones were placed in the water in both extraction groups which may be attributed to DNA loss and degradation. The findings contribute to the body of knowledge that informs DNA extraction procedures and methods used by the teams tasked with identifying victims of crime, natural disasters, and accidental deaths.

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