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Development of a microfluidic module for DNA purification via phenol extraction
Descriptive
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Title
Development of a microfluidic module for DNA purification via phenol extraction
Name
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Morales
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Mercedes C.
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Mercedes C. Morales
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author
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Zahn
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Jeffrey
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Advisory Committee
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Jeffrey D. Zahn
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chair
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Shinbrot
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Troy
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Advisory Committee
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Troy Shinbrot
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internal member
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Cai
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Li
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Advisory Committee
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Li Cai
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internal member
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Rutgers University
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degree grantor
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Graduate School - New Brunswick
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school
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theses
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2008
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2008-10
Language
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English
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electronic
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xi, 65 pages
Abstract
Purification of Deoxyribonucleic acid (DNA) by organic-aqueous liquid extraction, also called phenol extraction, is a standard technique commonly utilized in biology laboratories. In order to minimize interaction energies, membrane components and proteins naturally partition to the organic (phenol) phase while the DNA stays in the aqueous phase, where it can be easily removed. In recent years, microfluidics has become a driving force toward more efficient and autonomous platforms for fluid based diagnostics, chemical reaction chambers, separation and preparation of biological materials.
In this work, the design, fabrication, and performance of long microfluidic devices for DNA extraction are presented. The devices were fabricated using soft lithography to transfer lithographically defined features into a PDMS structure via replica molding. Stratified-flow experiments using a rhodamine dye conjugated bovine serum albumin protein (BSA) in an aqueous phase were conducted to compare different microchannel designs based on their ability to remove proteins from the aqueous phase into the phenol phase. Additionally, the study of BSA partitioning and DNA isolation in a two-phase system under stratified flow condition were addressed, separately and conjunctly. Finally, protein partitioning and DNA recovery were analyzed to evaluate two types of mixing, passive diffusion through stratified flows and droplet enhancement mixing.
In this work, the design, fabrication, and performance of long microfluidic devices for DNA extraction are presented. The devices were fabricated using soft lithography to transfer lithographically defined features into a PDMS structure via replica molding. Stratified-flow experiments using a rhodamine dye conjugated bovine serum albumin protein (BSA) in an aqueous phase were conducted to compare different microchannel designs based on their ability to remove proteins from the aqueous phase into the phenol phase. Additionally, the study of BSA partitioning and DNA isolation in a two-phase system under stratified flow condition were addressed, separately and conjunctly. Finally, protein partitioning and DNA recovery were analyzed to evaluate two types of mixing, passive diffusion through stratified flows and droplet enhancement mixing.
Note
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M.S.
Note
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Includes bibliographical references (p. 60-62).
Subject
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Topic
Biomedical Engineering
Subject
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Topic
DNA--Purification
Subject
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Topic
Microfluidic devices
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Title
Graduate School - New Brunswick Electronic Theses and Dissertations
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rucore19991600001
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http://hdl.rutgers.edu/1782.2/rucore10001600001.ETD.17533
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ETD_1146
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NjNbRU
Identifier
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doi:10.7282/T3T43TCX
Genre
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ETD graduate
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Rights
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The author owns the copyright to this work.
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Copyright protected
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Open
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Name
Mercedes Morales
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Copyright holder
Affiliation
Rutgers University. Graduate School - New Brunswick
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Non-exclusive ETD license
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Author Agreement License
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I hereby grant to the Rutgers University Libraries and to my school the non-exclusive right to archive, reproduce and distribute my thesis or dissertation, in whole or in part, and/or my abstract, in whole or in part, in and from an electronic format, subject to the release date subsequently stipulated in this submittal form and approved by my school. I represent and stipulate that the thesis or dissertation and its abstract are my original work, that they do not infringe or violate any rights of others, and that I make these grants as the sole owner of the rights to my thesis or dissertation and its abstract. I represent that I have obtained written permissions, when necessary, from the owner(s) of each third party copyrighted matter to be included in my thesis or dissertation and will supply copies of such upon request by my school. I acknowledge that RU ETD and my school will not distribute my thesis or dissertation or its abstract if, in their reasonable judgment, they believe all such rights have not been secured. I acknowledge that I retain ownership rights to the copyright of my work. I also retain the right to use all or part of this thesis or dissertation in future works, such as articles or books.
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