Molecular standards for analytical ultracentrifugation: investigating the suitability of double-stranded DNA

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dc.contributor.authorRanasinghe, Maduni Charuni
dc.contributor.authorUniversity of Lethbridge. Faculty of Arts and Science
dc.contributor.supervisorDemeler, Borries
dc.date.accessioned2023-11-30T22:29:23Z
dc.date.available2023-11-30T22:29:23Z
dc.date.issued2023
dc.degree.levelMasters
dc.description.abstractThis thesis focuses on the development of molecular standards to validate analytical ultracentrifugation (AUC) instruments, alongside an exploration of multiple applications of AUC. The goal of the main study is to address the critical need to develop a robust molecular standard for validating AUC instruments, with a specific emphasis on evaluating double-stranded DNA molecules as a potential candidate. By extensive investigation into the hydrodynamic properties of different topologies of double-stranded DNA at a wide range of temperatures, this study reveals the potential of linear and nicked double-stranded DNA as a reliable standard for AUC, contributing to the accurate characterization of macromolecules in solution. Supplementary research findings, as detailed in the attached publications presented in the attached appendix, further illustrate the versatility of AUC in various scientific domains. In one study, we employed AUC as a powerful technique to measure the sedimentation and diffusion coefficients of DNA minicircles to validate the elastic theory results obtained from our other collaborator. Also, it explores the impact of DNA supercoiling-induced shapes on minicircle hydrodynamics. In another collaborative effort, we study the oligomerization behavior of a de novo designed metalloprotein for photocatalytic hydrogen evolution, leveraging AUC for comprehensive characterization. These collective studies highlight the indispensable role of AUC in the characterization of macromolecules in solution, with applications ranging from DNA dynamics to functional protein characterization, and they demonstrate the crucial importance of reliable molecular standards in enhancing the accuracy of AUC measurements. Together, they contribute to the advancement of analytical science and its applications across diverse research domains.
dc.identifier.urihttps://hdl.handle.net/10133/6639
dc.language.isoen
dc.proquest.subject0487
dc.proquestyesYes
dc.publisherLethbridge, Alta. : University of Lethbridge, Dept. of Chemistry and Biochemistry
dc.publisher.departmentDepartment of Chemistry and Biochemistry
dc.publisher.facultyArts and Science
dc.relation.ispartofseriesThesis (University of Lethbridge. Faculty of Arts and Science)
dc.subjectanalytical ultracentrifugation
dc.subjectmolecular standards
dc.subjectdouble-stranded DNA
dc.subjectDNA minicircle
dc.subjecthydrodynamics
dc.subjectglobal analysis
dc.subject.lcshUltracentrifugation
dc.subject.lcshCentrifuges--Calibration--Research
dc.subject.lcshQuality control
dc.subject.lcshMacromolecules--Analysis
dc.subject.lcshNucleic acids
dc.subject.lcshSedimentation analysis
dc.subject.lcshGlobal analysis (Mathematics)
dc.subject.lcshDissertations, Academic
dc.titleMolecular standards for analytical ultracentrifugation: investigating the suitability of double-stranded DNA
dc.typeThesis
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