Functionalization of Magnetic Nanoparticles with Deoxynucleotide Triphosphates for Utilization in DNA Sequencing
2023-02
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Functionalization of Magnetic Nanoparticles with Deoxynucleotide Triphosphates for Utilization in DNA Sequencing
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2023-02
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Current methods for DNA sequencing leave a lot to be desired. There is immense research focused on developing methods to sequence longer and larger samples of DNA and to do so more rapidly. One possible new approach to DNA sequencing uses magnetically labeled deoxynucleotide triphosphates (dNTPs) as the complimentary bases for a sequencing-by-synthesis approach. The polymerase in this system would be immobilized on a sensor so that when the nucleotide is attached to the growing strand, the sensor can detect the signal. As the polymerase completes the strand for DNA extension, the signal from each magnetic nanoparticle (MNP) is recognized and generates a sequence.The contents of this thesis provide results for two critical initial steps in developing the proposed method. Firstly, we attached dNTPs to magnetic nanoparticles to create dNTP-MNP conjugates. We attempted two methods of conjugation with numerous characterization techniques to confirm the attachment. Secondly, we studied how the dNTP-MNP conjugation affected the dNTPs in quantitative polymerase chain reaction (qPCR) to model how they would interact with a polymerase in a hypothetical sequencer. The characterization results confirmed the conjugation of the dNTP to the MNP. qPCR was performed with various conditions and solvents which led to the conclusion that the MNP conjugation does not hinder polymerase interaction with the dNTP. This is a crucial result for the development of such a sequencer by providing the groundwork for the continuation of the project.
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University of Minnesota M.S.Ch.E. thesis. February 2023. Major: Chemical Engineering. Advisor: Kevin Dorfman. 1 computer file (PDF); viii, 50 pages.
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Nazareth, Calvin. (2023). Functionalization of Magnetic Nanoparticles with Deoxynucleotide Triphosphates for Utilization in DNA Sequencing. Retrieved from the University Digital Conservancy, https://hdl.handle.net/11299/256948.
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