자료유형  

 학위논문

Control Number  

0016930917

International Standard Book Number  

9798380583619

Dewey Decimal Classification Number  

574

Main Entry-Personal Name  

Hyman, Leland B.

Publication, Distribution, etc. (Imprint  

[S.l.] : The University of Wisconsin - Madison., 2020

Publication, Distribution, etc. (Imprint  

Ann Arbor : ProQuest Dissertations & Theses, 2020

Physical Description  

1 online resource(148 p.)

General Note  

Source: Dissertations Abstracts International, Volume: 85-04, Section: B.

General Note  

Advisor: Romero, Philip A.

Dissertation Note  

Thesis (Ph.D.)--The University of Wisconsin - Madison, 2020.

Restrictions on Access Note  

This item must not be sold to any third party vendors.

Summary, Etc.  

요약Intracellular heterogeneity affects the human body's response to drug treatment, pathogen infection, and disease processes. Methods to characterize single-cell transcriptional activities are currently expensive, laborious, and limited in cell throughput. By leveraging recent advances including droplet microfluidics, Loop-mediated Isothermal Amplification (LAMP), and DNA logic techniques, we have developed a simple, inexpensive, and high-throughput transcriptional profiling method: Single-cell Nucleic Acid Profiling in Droplets (SNAPD). We showed that this unprecedentedly simple workflow can analyze target gene expression in cell mixtures with incredible sensitivity and selectivity. We further enhanced SNAPD by developing a novel library of DNA logic gates which integrate with LAMP. We have demonstrated that SNAPD is an advantageous technique to study rare and mixed cell populations and could be powerful in many biological settings. We also applied these DNA logic principles to develop a novel method for accurate and reliable Single Nucleotide Polymorphism (SNP) detection. This method could be integrated with SNAPD for single-cell SNP assays, or applied directly to point-of-care diagnostics. In this work, we have developed two approaches which leverage LAMP and molecular biology techniques to create powerful research tools, proving their utility in oncological and diagnostic applications. Alone, each of these advances opens avenues for future work in diverse biological applications and diagnostics. Combined, they may comprise a single-cell SNP profiling method of unprecedented simplicity and performance.

Subject Added Entry-Topical Term  

Cellular biology.

Subject Added Entry-Topical Term  

Biomedical engineering.

Subject Added Entry-Topical Term  

Biochemistry.

Subject Added Entry-Topical Term  

Molecular biology.

Subject Added Entry-Topical Term  

Genetics.

Index Term-Uncontrolled  

Loop-mediated Isothermal Amplification

Index Term-Uncontrolled  

Microfluidics

Index Term-Uncontrolled  

Point-of-care diagnostics

Index Term-Uncontrolled  

Single-cell transcriptomics

Index Term-Uncontrolled  

DNA logic techniques

Added Entry-Corporate Name  

The University of Wisconsin - Madison Cellular & Molecular Bio - AG

Host Item Entry  

Dissertations Abstracts International. 85-04B.

Host Item Entry  

Dissertation Abstract International

Electronic Location and Access  

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Control Number  

joongbu:639878