Date of Award

5-2013

Level of Access Assigned by Author

Campus-Only Thesis

Degree Name

Master of Science (MS)

Department

Microbiology

Advisor

Paul Millard

Second Committee Member

John T. Singer

Third Committee Member

Rosemary Smith

Abstract

There is a critical and expanding need for effective control of explosives that can fall into the hands of those with malicious intent. An investigative tool that determined the origin and routes of transit of contraband explosive materials, through the use of taggants, would stem the theft and abuse of explosive materials. The focus of this thesis project was to generate a novel class of taggants and to fabricate a field-portable amplification and detection technology for on-site identification of these taggants.

Generation of the proposed taggant required bioengineering bacterial DNA to serve as a nucleic acid coded sequence for identifying a manufacturer and lot of explosive. Cloning of the sequence into the highly resistant endospores of Geobacillus thermoglucosidasius allows their unique DNA sequences to be protected from intentional or inadvertent destruction.

The fabrication of a disposable taggant amplification and detection device was carried out in polyimide silicone tape with subsequent bonding to microscope slide glass using the application of heat and a coronal discharge treatment. This composite device was designed to screen multiple taggant sequences in parallel using a modified quantitative polymerase chain reaction (qPCR) method. Detection of the unique DNA taggant sequences was carried out on-chip by measurement of start and end point fluorescence. In the composite device amplification of the bioengineered DNA sequence showed roughly a 9 times greater fluorescence signal over negative control samples.

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