Sequence specific colorimetric detection of DNA post-PCR using split DNAzyme

The split G-quadruplex DNAzyme has become a valuable tool for visual detection. Most reports on its use have however been constrained to proof-of-concept demonstrations using synthetic oligonucleotides. This is due to the inherent complexities of the assay, and the multiple components involved. H...

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Bibliographic Details
Main Author: Aw, Darius Kang Lie
Other Authors: Poh Chueh Loo
Format: Final Year Project (FYP)
Language:English
Published: 2010
Subjects:
Online Access:http://hdl.handle.net/10356/39817
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author Aw, Darius Kang Lie
author2 Poh Chueh Loo
author_facet Poh Chueh Loo
Aw, Darius Kang Lie
author_sort Aw, Darius Kang Lie
collection NTU
description The split G-quadruplex DNAzyme has become a valuable tool for visual detection. Most reports on its use have however been constrained to proof-of-concept demonstrations using synthetic oligonucleotides. This is due to the inherent complexities of the assay, and the multiple components involved. Herein, we have overcome several of these challenges and have successfully integrated asymmetric PCR with the visual detection step, allowing enriched targets from complex samples to be conveniently detected. This workflow brings the DNAzyme system closer to real-world detection applications. We have established the platform herein as a simple and robust method to determine the presence of target DNA sequences post-PCR, without the need for gels or other apparatus. Nanomolar concentrations of amplified targets were detected using the workflow established, enabling the detection of pathogenic targets and the discrimination of Single Nucleotide Polymorphism (SNP) targets through a color change reaction, observable just by eye.
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spelling ntu-10356/398172023-03-03T15:33:39Z Sequence specific colorimetric detection of DNA post-PCR using split DNAzyme Aw, Darius Kang Lie Poh Chueh Loo School of Chemical and Biomedical Engineering DSO National Laboratories Singapore DRNTU::Engineering::Chemical engineering::Biotechnology The split G-quadruplex DNAzyme has become a valuable tool for visual detection. Most reports on its use have however been constrained to proof-of-concept demonstrations using synthetic oligonucleotides. This is due to the inherent complexities of the assay, and the multiple components involved. Herein, we have overcome several of these challenges and have successfully integrated asymmetric PCR with the visual detection step, allowing enriched targets from complex samples to be conveniently detected. This workflow brings the DNAzyme system closer to real-world detection applications. We have established the platform herein as a simple and robust method to determine the presence of target DNA sequences post-PCR, without the need for gels or other apparatus. Nanomolar concentrations of amplified targets were detected using the workflow established, enabling the detection of pathogenic targets and the discrimination of Single Nucleotide Polymorphism (SNP) targets through a color change reaction, observable just by eye. Bachelor of Engineering (Chemical and Biomolecular Engineering) 2010-06-04T06:47:44Z 2010-06-04T06:47:44Z 2010 2010 Final Year Project (FYP) http://hdl.handle.net/10356/39817 en Nanyang Technological University 57 p. application/pdf
spellingShingle DRNTU::Engineering::Chemical engineering::Biotechnology
Aw, Darius Kang Lie
Sequence specific colorimetric detection of DNA post-PCR using split DNAzyme
title Sequence specific colorimetric detection of DNA post-PCR using split DNAzyme
title_full Sequence specific colorimetric detection of DNA post-PCR using split DNAzyme
title_fullStr Sequence specific colorimetric detection of DNA post-PCR using split DNAzyme
title_full_unstemmed Sequence specific colorimetric detection of DNA post-PCR using split DNAzyme
title_short Sequence specific colorimetric detection of DNA post-PCR using split DNAzyme
title_sort sequence specific colorimetric detection of dna post pcr using split dnazyme
topic DRNTU::Engineering::Chemical engineering::Biotechnology
url http://hdl.handle.net/10356/39817
work_keys_str_mv AT awdariuskanglie sequencespecificcolorimetricdetectionofdnapostpcrusingsplitdnazyme