Date of Award

2009

Degree Type

Dissertation

Department

Chemistry

First Advisor

Bayachou, Mekki

Subject Headings

DNA damage, Biosensors, Chemistry, Analytic, DNA mismatch detection, AFM, DNA damage detection, electrochemical sensors

Abstract

A large number of genetic diseases and genetic disorders are simply caused by base alterations in the genome. Therefore, developing efficient and cost effective techniques for routine detection of these alterations is of great importance. Different methods involving gel electrophoresis and Polymerase Chain Reaction have been widely employed, but majority of these methods are costly, time consuming, and lack throughput, creating a fundamental gap between the current state-of-the-art and desired characteristics of low-cost, high-speed, simplicity, versatility, and potential for miniaturization. In this study, we attempt to bridge this gap by developing new sensing platforms to detect DNA base mismatches and DNA damage with higher throughput, better ease-of-use, and with the potential to be miniaturized for greater portability. Two electrochemical mismatch detection sensing platforms were developed. One uses the electrochemical reduction of trans-4-cinnamic acid diazonium tetrafluoroborate. The other takes advantage of the natural ability of MutS protein for single base mismatch recognition. Also, two DNA damage detection assays were developed and the first approach uses Atomic Force Microscopy to monitor minor DNA damage by labeling damaged sites with a biomarker. This site-specific biolabeling was achieved through well-established biotin-streptavidin chemistry. In the second approach, a new layer-by-layer biomolecular immobilization method was introduced and used to detect DNA chemical damage using electrochemical techniques

Download

Included in

Chemistry Commons

COinS