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DEVELOPMENTS TOWARDS SINGLE MOLECULE AND SINGLE CELL GENETIC AND EPIGENETIC ANALYSIS TECHNOLOGIES

Author
Tian, Harvey Chen
Abstract
New discoveries in the realm of biology often come hand-in-hand with innovations in technology. Over the last two decades, advancements in microfabrication and microscopy have opened biological phenomena to be studied in fundamental units rather than as a collective entity. From these efforts, the axioms upon which we built our fundamental concepts in anatomy and physiology have been reshaped. We now understand that even within a subpopulation of cells taken from the same tissue, cell-to-cell variations exist in both phenotypic and genotypic expression. These variations are found to play critical roles in disease state and progression, thus, research into single cells and single molecules are a necessity to improving treatment.
In this work, methods of single molecule analysis of native human chromatin fibers are presented as well as methods and devices for multi- and single cell genome analysis. As the handling of single molecules and single cells are facilitated by microfabricated devices, discussed herein are their designs, fabrication methods, operational procedures, working principles, and experimental results representative of their biologically relevant impact. Our findings point towards potential platform technologies in high-throughput chromatin linearization for fluorescence based epigenetic mapping and single cell whole genome amplification with reduced amplification bias and improved
genome coverage.
Date Issued
2017-05-30Subject
Amplification; DNA; Microfluidic; Micropillar; Sequencing; Single Cell; Genetics; Biomedical engineering
Committee Chair
Craighead, Harold G
Committee Member
Luo, Dan; Zipfel, Warren R
Degree Discipline
Biomedical Engineering
Degree Name
Ph. D., Biomedical Engineering
Degree Level
Doctor of Philosophy
Type
dissertation or thesis