Date of Award

Summer 8-2017

Level of Access Assigned by Author

Campus-Only Thesis

Degree Name

Master of Science (MS)

Department

Biological Engineering

Advisor

Paul Millard

Second Committee Member

Mauricio Pereira da Cunha

Third Committee Member

John Singer

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. A proposed, novel method of taggant generation involves the creation of bioengineered bacteria that contains specific DNA taggant sequences. The focus of this thesis project was the development and fabrication of a lysis device that would allow for field-portable extraction of DNA from modified vegetative bacteria for on-site identification of these taggants.

Lysis of the bacteria, Geobacillus thermoglucosidasius, required the development of a surface acoustic wave (SAW) device. SAW technology was employed for its ability to be miniaturized, use low amounts of power, and the potential to provide high efficiency for cell lysis due to SAW energy trapping at the surface of the device.

The fabrication of the SAW device was carried out on Y-128° lithium niobate (LNO). Operating at 96 MHz and held at surface temperatures of 5°, 35°, and 55°C, the SAW device successfully lysed vegetative bacteria. DNA released through lysis was quantified using a PicoGreen fluorescent stain and revealed that up to 30% of all cells were lysed after 240 seconds of exposure at 35°C. Quantitative polymerase chain reaction (qPCR) was performed to confirm that DNA released originated from Geobacillus thermoglucosidasius. Further testing on a different cell type, bovine red blood cells, showed that the system could be used to lyse a different cell type, leading to potential uses outside of a bacterial taggant detection system.

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