DNA digestion and ligation on a microfluidic chip for rapid gene engineering

A microfluidic system for fluorescence detection on digested and ligated DNA fragments was developed as one of the development for rapid genetic engineering. This system was integrated onto a silicon chip with enzymatic reaction taken place within the functionalized chip thereafter. Functionalized c...

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Main Author: Tan, Sharon Xue Ni.
Other Authors: Chan Chi Chiu
Format: Final Year Project
Language:English
Published: 2011
Subjects:
Online Access:http://hdl.handle.net/10356/45169
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Institution: Nanyang Technological University
Language: English
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spelling sg-ntu-dr.10356-451692023-03-03T15:37:36Z DNA digestion and ligation on a microfluidic chip for rapid gene engineering Tan, Sharon Xue Ni. Chan Chi Chiu School of Chemical and Biomedical Engineering A*STAR Institute of Microelectronics DRNTU::Engineering::Chemical engineering::Biotechnology A microfluidic system for fluorescence detection on digested and ligated DNA fragments was developed as one of the development for rapid genetic engineering. This system was integrated onto a silicon chip with enzymatic reaction taken place within the functionalized chip thereafter. Functionalized chip technique was a key process for better binding affinity between the chip surface and the DNA. By making use of fluorescent dye, measurement of splicing and attachments segments was possible. This technique will allow a DNA restriction map to be construe using an automated system by optical mapping quickly.Stepwise functionalizing of chip with APTES, biotin and avidin were completed within 4 hours. Biotin-synthesize double stranded DNA (dsDNA) labeled with FAM and CY5 at each ends respectively was immobilized onto the surface of the chip within 1 hour. Digestion by BamH1 and ligation by DNA T4 ligase were both favorable at 5 minutes. Compared this micro-system to conventional molecular diagnostic, which will took 4 hours for digestion and 12 hours for ligation, such system had its advantages over time and sample volume containment. Bachelor of Engineering (Chemical and Biomolecular Engineering) 2011-06-09T07:32:48Z 2011-06-09T07:32:48Z 2011 2011 Final Year Project (FYP) http://hdl.handle.net/10356/45169 en Nanyang Technological University 37 p. application/pdf
institution Nanyang Technological University
building NTU Library
continent Asia
country Singapore
Singapore
content_provider NTU Library
collection DR-NTU
language English
topic DRNTU::Engineering::Chemical engineering::Biotechnology
spellingShingle DRNTU::Engineering::Chemical engineering::Biotechnology
Tan, Sharon Xue Ni.
DNA digestion and ligation on a microfluidic chip for rapid gene engineering
description A microfluidic system for fluorescence detection on digested and ligated DNA fragments was developed as one of the development for rapid genetic engineering. This system was integrated onto a silicon chip with enzymatic reaction taken place within the functionalized chip thereafter. Functionalized chip technique was a key process for better binding affinity between the chip surface and the DNA. By making use of fluorescent dye, measurement of splicing and attachments segments was possible. This technique will allow a DNA restriction map to be construe using an automated system by optical mapping quickly.Stepwise functionalizing of chip with APTES, biotin and avidin were completed within 4 hours. Biotin-synthesize double stranded DNA (dsDNA) labeled with FAM and CY5 at each ends respectively was immobilized onto the surface of the chip within 1 hour. Digestion by BamH1 and ligation by DNA T4 ligase were both favorable at 5 minutes. Compared this micro-system to conventional molecular diagnostic, which will took 4 hours for digestion and 12 hours for ligation, such system had its advantages over time and sample volume containment.
author2 Chan Chi Chiu
author_facet Chan Chi Chiu
Tan, Sharon Xue Ni.
format Final Year Project
author Tan, Sharon Xue Ni.
author_sort Tan, Sharon Xue Ni.
title DNA digestion and ligation on a microfluidic chip for rapid gene engineering
title_short DNA digestion and ligation on a microfluidic chip for rapid gene engineering
title_full DNA digestion and ligation on a microfluidic chip for rapid gene engineering
title_fullStr DNA digestion and ligation on a microfluidic chip for rapid gene engineering
title_full_unstemmed DNA digestion and ligation on a microfluidic chip for rapid gene engineering
title_sort dna digestion and ligation on a microfluidic chip for rapid gene engineering
publishDate 2011
url http://hdl.handle.net/10356/45169
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