University of Denver
Sequence Detection and Comparative Analysis of the Hv1 and Hv2 Control Regions of Human Mitochondrial DNA by Denaturing High-Performance Liquid Chromatography
Abstract
dc:description.abstract<p>The objective of this research was the evaluation and forensic validation of Denaturing High-Performance Liquid Chromatography (DHPLC) as a sequencingindependent means of detecting the presence of sequence differences in pair-wise mixtures of non-concordant amplicons of human mitochondrial DNA (mtDNA). The reproducibility and efficacy of DHPLC results, including amplification reproducibility, injection reproducibility, and column-to-column reproducibility were measured, showing negligible assay-to-assay variability. In addition, cross-contamination on the DHPLC columns demonstrated very low level DNA carryover between a high-abundance sample and subsequent zero-volume injections.</p> <p>The accuracy with which DHPLC technology can be used to screen both evidence and control samples in the context of a forensic laboratory was evaluated. This was demonstrated by a number of pair-wise comparisons of each of the forensically relevant amplicons from 95 unrelated individuals in the study, and was in 100% agreement with sequencing data. Thus, DHPLC can be used to detect a diversity of sequence differences (transitions, transversions, insertions and deletions) in the mtDNA D-loop. Accordingly, DHPLC may have utility as a presumptive indicator of mtDNA sequence concordance samples.</p>
Degree
thesis:*- Name thesis:degree_name
- M.S.
- Level thesis:degree_level
- Masters Thesis
- Year dc:date.available
- 2011
Author and committee
dc:creator, dc:contributor.*- Author dc:creator
-
- Lewis, Sarah E.
- Contributors dc:contributor
-
- Phillip B. Danielson, Ph.D.
- James Fogleman
- Nancy Lorenzon
- Davor Balzar
Subjects
dc:subject × 5Rights
dc:rights- Statement dc:rights
-
- <p>Copyright is held by the author. User is responsible for all copyright compliance.</p>
- Language dc:language
- en
Identifiers
dc:identifier.*- Repository record dc:identifier
- https://digitalcommons.du.edu/etd/365
- OAI identifier oai:identifier
- oai:digitalcommons.du.edu:etd-1364