Date of Submission
5-2025
Document Type
Thesis
Degree Name
Master of Science in Forensic Science
Department
Forensic Science
Advisor
Ashley G. Morgan, Ph.D.
Committee Member
Heather M. Coyle, Ph.D.
Committee Member
Sharon N. Kahara, PhD.
Keywords
DNA degradation, Skeletal Remains Submerged in Stagnant Water, Forensic DNA Extraction, Prepfiler BTA Protocol, Proteinase K
MeSH
DNA, Bone and Bones, Body Remains
LCSH
DNA—Analysis, Human decomposition
Abstract
Successful genetic identification of human remains is heavily dependent on environmental factors. This sentiment is stressed when considering those submerged in aqueous environments. However, the range of studies investigating the interaction of DNA degradation and skeletal remains is quite limited - especially those submerged in water. Consequently, this study aimed to quantify and determine the degradation rate of submerged skeletal element DNA in stagnant, tannin-rich blackwater wetland over a four-month period. To mimic human remains in a New England Pond environment, five macerated porcine femurs were submerged in an artificial pond aquarium and allowed to soak for three weeks, six weeks, nine weeks, 12 weeks, and 15 weeks respectively. This aquarium began as an acidic, tannin-rich, and stagnant body of water, that supported plant life. A control sample was left in dry, ambient room conditions and was analyzed in tandem with the submerged samples at the end of each period. Bone samples were cut, dried, then powdered, and DNA was extracted using the Prepfiler™ BTA Forensic DNA Extraction Kit. The following two methods were used for this extraction: the Applied Biosystems™ recommended Prepfiler™ BTA protocol and its updated protocol. Following extraction, samples were simultaneously amplified and quantified using the QuantStudio TM 5 Real-Time PCR System. No experimental samples amplified in the first nine weeks, but three amplified to low template levels(< 0.04ng/µL) at 12 and 15 weeks. Nearly all the dry control samples yielded DNA (~93%). Of those control samples, the recommended Prepfiler™ BTA protocol produced higher DNA yields than the updated protocol at all measurement periods. This suggests that the inclusion of Proteinase K during extraction correlates to higher DNA yield. Consequently, forensic laboratories should prioritize this method in skeletal DNA recovery efforts.
Recommended Citation
Behn, Jessica R., "Evaluation of Skeletal DNA From Bones Submerged in Stagnant, Tannin-Rich Blackwater" (2025). Master's Theses. 241.
https://digitalcommons.newhaven.edu/masterstheses/241