High-throughput genetic analysis and combinatorial chiral separations based on capillary electrophoresis

dc.contributor.advisor Edward S. Yeung
dc.contributor.author Zhong, Wenwan
dc.contributor.department Chemistry
dc.date 2018-08-25T01:10:37.000
dc.date.accessioned 2020-07-02T06:01:28Z
dc.date.available 2020-07-02T06:01:28Z
dc.date.copyright Wed Jan 01 00:00:00 UTC 2003
dc.date.issued 2003-01-01
dc.description.abstract <p>Capillary electrophoresis offers many advantages over conventional analytical methods, such as speed, simplicity, high resolution, low cost, and small sample consumption, especially for the separation of enantiomers. However, chiral method developments still can be time consuming and tedious. We designed a comprehensive enantioseparation protocol employing neutral and sulfated cyclodextrins as chiral selectors for common basic, neutral, and acidic compounds with a 96-capillary array system. By using only four judiciously chosen separation buffers, successful enantioseparations were achieved for 49 out of 54 test compounds spanning a large variety of pKs and structures. Therefore, unknown compounds can be screened in this manner to identify optimal enantioselective conditions in just one run. In addition to superior separation efficiency for small molecules, CE is also the most powerful technique for DNA separations. Using the same multiplexed capillary system with UV absorption detection, DNA sequencing of a short template was done without any dye-labels. Two internal standards were utilized to adjust the migration time variations among capillaries, so that the four electropherograms for the A, T, C, G Sanger reactions can be aligned and base calling can be completed with a level of high confidence. The CE separation of DNA can be applied to study differential gene expression as well. Combined with pattern recognition techniques, small variations among electropherograms obtained by the separation of cDNA fragments produced from the total RNA samples of different human tissues can be revealed. These variations reflect the differences in total RNA expression among tissues. Thus, this CE-based approach can serve as an alternative to the DNA array techniques in gene expression analysis.</p>
dc.format.mimetype application/pdf
dc.identifier archive/lib.dr.iastate.edu/rtd/759/
dc.identifier.articleid 1758
dc.identifier.contextkey 6080496
dc.identifier.doi https://doi.org/10.31274/rtd-180813-12119
dc.identifier.s3bucket isulib-bepress-aws-west
dc.identifier.submissionpath rtd/759
dc.identifier.uri https://dr.lib.iastate.edu/handle/20.500.12876/80483
dc.language.iso en
dc.source.bitstream archive/lib.dr.iastate.edu/rtd/759/r_3118273.pdf|||Sat Jan 15 01:50:23 UTC 2022
dc.subject.disciplines Analytical Chemistry
dc.subject.keywords Chemistry
dc.subject.keywords Analytical chemistry
dc.title High-throughput genetic analysis and combinatorial chiral separations based on capillary electrophoresis
dc.type article
dc.type.genre dissertation
dspace.entity.type Publication
relation.isOrgUnitOfPublication 42864f6e-7a3d-4be3-8b5a-0ae3c3830a11
thesis.degree.level dissertation
thesis.degree.name Doctor of Philosophy
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