In situ formed magnetic ionic liquids: DNA extraction performance and fluorescence-compatibility in bioanalytical applications

dc.contributor.advisor Jared L. Anderson Bowers, Ashley
dc.contributor.department Chemistry 2020-02-12T22:53:18.000 2020-06-30T03:19:55Z 2020-06-30T03:19:55Z Sun Dec 01 00:00:00 UTC 2019 2020-05-23 2019-01-01
dc.description.abstract <p>Extracting genomic DNA from complex biological sample matrices is often the first step in numerous molecular biology procedures such as polymerase chain reaction (PCR), cloning, and gene therapy. Obtaining high yields and pure DNA presents a significant sample preparation challenge in nucleic acid analysis. Current methodologies such as the phenol-chloroform extraction use toxic organic solvents and commercially available kits are often very expensive and have limited reusability. Magnetic ionic liquids (MILs) have gained popularity as inexpensive, environmentally benign and tunable extraction solvents. MILs are a subclass of ionic liquids containing a paramagnetic component in the cation or anion, allowing them to be manipulated using an external magnetic field. This thesis describes the use of a new class of MILs featuring metal-containing cations for DNA extraction and their compatibility with fluorescence-based detection methods. Two studies were conducted to address this goal. The first study focused on the DNA extraction efficiency of a new class of MILs using in situ dispersive liquid-liquid microextraction (DLLME) versus conventional DLLME to assess the extraction of DNA sequences of varied sizes. Extraction efficiencies were obtained using indirect detection using anion-exchange high performance liquid chromatography with diode array detection and fluorescence spectroscopy. However, to minimize steps in the sample preparation process, it is useful to directly analyze the DNA within the enriched MIL microdroplet therefore, in a second study the fluorescence quenching effects of the MIL were evaluated. These studies provide an insight into how the paramagnetic metal (Ni, Co, Mn) and ligand used in the design of the MIL can be tailored in order to achieve highly efficient DNA extraction and the subsequent influence of the MIL on the fluorescence signal in downstream analysis.</p>
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dc.identifier archive/
dc.identifier.articleid 8655
dc.identifier.contextkey 16524485
dc.identifier.s3bucket isulib-bepress-aws-west
dc.identifier.submissionpath etd/17648
dc.language.iso en
dc.source.bitstream archive/|||Fri Jan 14 21:26:46 UTC 2022
dc.subject.disciplines Analytical Chemistry
dc.subject.disciplines Chemistry
dc.subject.disciplines Physical Chemistry
dc.subject.keywords DNA
dc.subject.keywords fluorescence spectroscopy
dc.subject.keywords high performance liquid chromatography
dc.subject.keywords in situ dispersive liquid-liquid microextraction
dc.subject.keywords magnetic ionic liquids
dc.subject.keywords SYBR Green I
dc.title In situ formed magnetic ionic liquids: DNA extraction performance and fluorescence-compatibility in bioanalytical applications
dc.type article
dc.type.genre thesis
dspace.entity.type Publication
relation.isOrgUnitOfPublication 42864f6e-7a3d-4be3-8b5a-0ae3c3830a11 Analytical Chemistry thesis Master of Science
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