Development and implementation of a T-RFLP procedure for use on bioreactor sludge

Terminal restriction fragment length polymorphism analysis is popularly used as fast and reliable method to investigate the profile of microbial communities in environmental samples. It can be used as a fingerprinting technique to investigate the structure and composition of bacteria that are presen...

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Main Author: Tan, Daniel Chiao Thien
Other Authors: School of Civil and Environmental Engineering
Format: Final Year Project
Language:English
Published: 2013
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Online Access:http://hdl.handle.net/10356/53904
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Institution: Nanyang Technological University
Language: English
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spelling sg-ntu-dr.10356-539042023-03-03T17:23:40Z Development and implementation of a T-RFLP procedure for use on bioreactor sludge Tan, Daniel Chiao Thien School of Civil and Environmental Engineering Singapore Centre for Environmental Life Sciences Engineering Stefan Wuertz DRNTU::Engineering Terminal restriction fragment length polymorphism analysis is popularly used as fast and reliable method to investigate the profile of microbial communities in environmental samples. It can be used as a fingerprinting technique to investigate the structure and composition of bacteria that are present in the wastewater treatment processes. In this project, I developed a T-RFLP method for wastewater sludge and used this method to investigate the bacterial community in sludge from four membrane bioreactors. DNA extracted from bioreactor sludge samples underwent Polymerase Chain Reaction to amplify the 16S rRNA gene; this amplicon then underwent PCR product purification, and the purified amplicon was finally enzyme digestion whereby a variety of enzymes were used to obtain Terminal-Restriction Fragments. A major portion of the project involved optimization of these steps in the T-RFLP pipeline. Data analysis was then done to identify which enzyme gave the most number of true peaks and highest fluorescence. This T-RFLP pipeline was used to examine the microbial diversity of sludge in four MBRs and the communities were compared quantitatively using three diversity indices, namely richness, dynamics and evenness. The optimized procedure developed in my project will also continue to be used as the standard T-RFLP protocol in the Experimental Bioprocess Evolution group at Singapore Centre of Environmental Life Sciences Engineering (SCELSE). Bachelor of Engineering (Environmental Engineering) 2013-06-10T03:38:44Z 2013-06-10T03:38:44Z 2013 2013 Final Year Project (FYP) http://hdl.handle.net/10356/53904 en Nanyang Technological University 41 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
spellingShingle DRNTU::Engineering
Tan, Daniel Chiao Thien
Development and implementation of a T-RFLP procedure for use on bioreactor sludge
description Terminal restriction fragment length polymorphism analysis is popularly used as fast and reliable method to investigate the profile of microbial communities in environmental samples. It can be used as a fingerprinting technique to investigate the structure and composition of bacteria that are present in the wastewater treatment processes. In this project, I developed a T-RFLP method for wastewater sludge and used this method to investigate the bacterial community in sludge from four membrane bioreactors. DNA extracted from bioreactor sludge samples underwent Polymerase Chain Reaction to amplify the 16S rRNA gene; this amplicon then underwent PCR product purification, and the purified amplicon was finally enzyme digestion whereby a variety of enzymes were used to obtain Terminal-Restriction Fragments. A major portion of the project involved optimization of these steps in the T-RFLP pipeline. Data analysis was then done to identify which enzyme gave the most number of true peaks and highest fluorescence. This T-RFLP pipeline was used to examine the microbial diversity of sludge in four MBRs and the communities were compared quantitatively using three diversity indices, namely richness, dynamics and evenness. The optimized procedure developed in my project will also continue to be used as the standard T-RFLP protocol in the Experimental Bioprocess Evolution group at Singapore Centre of Environmental Life Sciences Engineering (SCELSE).
author2 School of Civil and Environmental Engineering
author_facet School of Civil and Environmental Engineering
Tan, Daniel Chiao Thien
format Final Year Project
author Tan, Daniel Chiao Thien
author_sort Tan, Daniel Chiao Thien
title Development and implementation of a T-RFLP procedure for use on bioreactor sludge
title_short Development and implementation of a T-RFLP procedure for use on bioreactor sludge
title_full Development and implementation of a T-RFLP procedure for use on bioreactor sludge
title_fullStr Development and implementation of a T-RFLP procedure for use on bioreactor sludge
title_full_unstemmed Development and implementation of a T-RFLP procedure for use on bioreactor sludge
title_sort development and implementation of a t-rflp procedure for use on bioreactor sludge
publishDate 2013
url http://hdl.handle.net/10356/53904
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