Development of clean up protocol for sensitive DNA colorimetric assays

An analytical Deoxyribo Nucleic Acid (DNA) colorimetric assay was done to find out how the detection limit are impinged in the presence of Bovine Serum albumin (BSA) in solution. DNA is known to be used as potential biomarkers for diagnosis of major diseases, such as cancer and heart diseases. Howev...

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Main Author: Kwek, Tian Chen
Other Authors: Bo Liedberg
Format: Final Year Project (FYP)
Language:English
Published: Nanyang Technological University 2020
Subjects:
Online Access:https://hdl.handle.net/10356/139164
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author Kwek, Tian Chen
author2 Bo Liedberg
author_facet Bo Liedberg
Kwek, Tian Chen
author_sort Kwek, Tian Chen
collection NTU
description An analytical Deoxyribo Nucleic Acid (DNA) colorimetric assay was done to find out how the detection limit are impinged in the presence of Bovine Serum albumin (BSA) in solution. DNA is known to be used as potential biomarkers for diagnosis of major diseases, such as cancer and heart diseases. However, the presence of interferents such as BSA affect the sensitivity of such biomarkers. While there are many detection techniques for DNA, there are often expensive, complex and require trained personnel. Thus, in this study, we made use of the optical properties conjugated polymers (CPs) to clean up BSA to improve sensitivity. In addition, CPs is used to detect DNA because of DNA’s fluorescence quenching abilities. The binding of DNA to cationic conjugated polymer through electrostatic interactions and docking of BSA to anionic polythiophene has been reported earlier. Two readily available conjugated polymers namely cationic polythiophene-1 (PT1) and anionic polythiophene-4(PT4) were used to demonstrate the fluorescence quenching after adding the biomolecules. Each polymer was added to react individually with DNA solutions, BSA solutions and a mixture of DNA and BSA solutions, respectively. Several parameters were varied, including volume, concentration of PTs and the different buffers for optimization. The absorbance and fluorescence spectra of the resulting solutions were measured using UV-Visible and fluorescence spectroscopy, respectively.
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spelling ntu-10356/1391642023-03-04T15:48:21Z Development of clean up protocol for sensitive DNA colorimetric assays Kwek, Tian Chen Bo Liedberg School of Materials Science and Engineering bliedberg@ntu.edu.sg Engineering::Materials An analytical Deoxyribo Nucleic Acid (DNA) colorimetric assay was done to find out how the detection limit are impinged in the presence of Bovine Serum albumin (BSA) in solution. DNA is known to be used as potential biomarkers for diagnosis of major diseases, such as cancer and heart diseases. However, the presence of interferents such as BSA affect the sensitivity of such biomarkers. While there are many detection techniques for DNA, there are often expensive, complex and require trained personnel. Thus, in this study, we made use of the optical properties conjugated polymers (CPs) to clean up BSA to improve sensitivity. In addition, CPs is used to detect DNA because of DNA’s fluorescence quenching abilities. The binding of DNA to cationic conjugated polymer through electrostatic interactions and docking of BSA to anionic polythiophene has been reported earlier. Two readily available conjugated polymers namely cationic polythiophene-1 (PT1) and anionic polythiophene-4(PT4) were used to demonstrate the fluorescence quenching after adding the biomolecules. Each polymer was added to react individually with DNA solutions, BSA solutions and a mixture of DNA and BSA solutions, respectively. Several parameters were varied, including volume, concentration of PTs and the different buffers for optimization. The absorbance and fluorescence spectra of the resulting solutions were measured using UV-Visible and fluorescence spectroscopy, respectively. Bachelor of Engineering (Materials Engineering) 2020-05-16T13:21:50Z 2020-05-16T13:21:50Z 2020 Final Year Project (FYP) https://hdl.handle.net/10356/139164 en application/pdf Nanyang Technological University
spellingShingle Engineering::Materials
Kwek, Tian Chen
Development of clean up protocol for sensitive DNA colorimetric assays
title Development of clean up protocol for sensitive DNA colorimetric assays
title_full Development of clean up protocol for sensitive DNA colorimetric assays
title_fullStr Development of clean up protocol for sensitive DNA colorimetric assays
title_full_unstemmed Development of clean up protocol for sensitive DNA colorimetric assays
title_short Development of clean up protocol for sensitive DNA colorimetric assays
title_sort development of clean up protocol for sensitive dna colorimetric assays
topic Engineering::Materials
url https://hdl.handle.net/10356/139164
work_keys_str_mv AT kwektianchen developmentofcleanupprotocolforsensitivednacolorimetricassays