The Development of a Custom qPCR Multiplex to Quantify Human and Microbial DNA
Open Access
Author:
Foster, Miriam
Area of Honors:
Forensic Science
Degree:
Bachelor of Science
Document Type:
Thesis
Thesis Supervisors:
Mitchell Mark Holland, Thesis Supervisor Mitchell Mark Holland, Thesis Honors Advisor Jason Brooks, Faculty Reader
Keywords:
DNA Forensic Biology Forensic Microbiology qPCR
Abstract:
Past work researching microbial forensics has focused solely on sequencing and has neglected the reporting and expectations of the quantification of, as well as the comparison between, human and bacterial DNA. The goal of this work was to produce a novel multiplex qPCR assay that allows for the simultaneous quantification of human and bacterial DNA. Through the use of this assay, expectations for the amounts of each type of DNA, human and bacterial, that could be found on certain surfaces or sample types were produced. Through repeatability studies and several small-scale experiments, the viability of this assay was established. For example, practical expectations of DNA quantities were found for items such as keyboards, computer mice, cell phones, smart watches, and others. The qPCR multiplex and information provided by this work will help analysts hoping to identify and connect individuals decide whether they should pursue human or bacterial DNA analysis methods, depending on the available quantities of each.