Date of Award

2013

Document Type

Dissertation

Degree Name

Doctor of Philosophy (PhD)

Department

Biotechnology Science and Engineering

Committee Chair

Roy Magnuson

Committee Member

Debra M. Moriarity

Committee Member

Joseph Ng

Committee Member

Marshall Schreeder

Committee Member

Jian Han

Subject(s)

Polymerase chain reaction, RNA microarrays, Medical screening, Cancer--Diagnosis, RNA--Analysis, Gene expression

Abstract

Ligation-dependent PCR, when coupled with a bead-based detection array, comprises a method of nucleic acid detection that utilizes enzymes and equipment already being employed successfully in clinical labs. The process can be multiplexed in order to generate a profile of the sequences of interest. When applied to microRNA profiling, the ligation step requires modification to improve the efficiency of the T4 DNA ligase employed. With optimization, the assay has the potential to distinguish and detect multiple microRNA species simultaneously, providing a clinically adaptable method for microRNA profiling. The flexibility of the assay format would allow modification for a nearly limitless repertoire of disease-specific panels.

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