Defense Date
2023
Document Type
Directed Research Project
First Advisor
Tal Simmons, Ph.D.
Second Advisor
Baneshwar Singh, Ph.D.
Third Advisor
Joseph Jones, Ph.D.
Abstract
During the construction on the Virginia Commonwealth University medical campus in 1994, human remains and artifacts were discovered in what is now referred to as the East Marshall Street Well (EMSW). Artifacts found in the well can be dated back to the 19th century and human remains discovered were likely medical cadavers obtained illegally back in mid-19th century, when grave robbing was widely practiced. As a result, the East Marshall Street Well Project (EMSWP) was established in 2015, following the raising of public consciousness concerning the discovery. The aim of this study is to verify previously pair-matched arm element groups (performed by anthropologists at the Smithsonian using osteometric analysis) using Insertion/Null (INNULs) genotyping in order to assemble individuals for reburial purposes. Due to the bones recovered from EMSW being highly degraded, an optimization study was also performed prior to genotyping in order to determine which method was best suited to extract quality DNA from the remains. In total, three sampling methods (scalpel scraping, Dremel sanding, and cut bone sampling) in combination with three extraction methods (organic phenol-chloroform extraction, a modified organic phenol-chloroform extraction suggested by Loreille et al. (2007), and InnoXtract™️ Bone Extraction Kit) were tested on four femurs of donated individuals obtained for teaching purposes. Quantitation of DNA was performed using the InnoQuantÒ HY kit on QuantStudio™️ 6 Real-time PCR. Even though no statistical significance was found between the different combinations of sampling and extraction methods, the combination of the Dremel sampling method and InnoXtract™️ Bone Extraction Kit resulted in the second highest DNA yield and lowest degradation index. Therefore, the combination of the Dremel sampling and InnoXtract™️ Bone Extraction Kit was used to extract DNA from EMSW remains. The quantitation step was performed using the InnoQuantÒ HY kit and genotyping was performed using the InnoTyperⓇ 21 Human DNA Typing kit. Of 106 bone samples, 52 yielded full or partial profiles suitable for comparison, and 41 of them were included in the percent match calculation. Of the 41 bones, 28 (68.29%) bones were correctly matched within the group, 7 (17.07%) bones cannot be excluded as a match, and 6 (14.63%) bones were incorrectly matched. As suggested from the DNA extraction optimization portion of the study, Dremel sanding sampling method could be an alternative to the cutbone sampling method when there is a need to preserve the integrity of the bones. Overall, more than half of the bones were previously pair-matched correctly with some bones that were incorrectly matched, indicating that pair-matching could be a useful method for estimating bone groups but should ultimately be verified by DNA analysis.
Rights
© The Author(s)
Is Part Of
VCU Master of Science in Forensic Science Directed Research Projects
Date of Submission
5-5-2023