Date of Award

8-1-2011

Document Type

Thesis

Degree Name

M.S.

Department

Biological Sciences

First Advisor

Phillip B. Danielson, Ph.D.

Abstract

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.

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.

Provenance

Received from ProQuest

Rights holder

Sarah E. Lewis

File size

102 p.

File format

application/pdf

Language

en

Discipline

Biology

Included in

Biology Commons

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