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Title: Protocol Improvements for Low Concentration DNA-Based Bioaerosol Sampling and Analysis
Authors: Luhung, Irvan
Wu, Yan
Ng, Chun Kiat
Miller, Dana
Cao, Bin
Chang, Victor Wei-Chung
Keywords: Civil and Environmental Engineering
Issue Date: 2015
Source: Luhung, I., Wu, Y., Ng, C. K., Miller, D., Cao, B.,& Chang, V. W. C. (2015). Protocol Improvements for Low Concentration DNA-Based Bioaerosol Sampling and Analysis. PLOS ONE, 10(11), e0141158-.
Series/Report no.: PLOS ONE
Abstract: Introduction: As bioaerosol research attracts increasing attention, there is a need for additional efforts that focus on method development to deal with different environmental samples. Bioaerosol environmental samples typically have very low biomass concentrations in the air, which often leaves researchers with limited options in choosing the downstream analysis steps, especially when culture-independent methods are intended. Objectives: This study investigates the impacts of three important factors that can influence the performance of culture-independent DNA-based analysis in dealing with bioaerosol environmental samples engaged in this study. The factors are: 1) enhanced high temperature sonication during DNA extraction; 2) effect of sampling duration on DNA recoverability; and 3) an alternative method for concentrating composite samples. In this study, DNA extracted from samples was analysed using the Qubit fluorometer (for direct total DNA measurement) and quantitative polymerase chain reaction (qPCR). Results and Findings: The findings suggest that additional lysis from high temperature sonication is crucial: DNA yields from both high and low biomass samples increased up to 600% when the protocol included 30-min sonication at 65°C. Long air sampling duration on a filter media was shown to have a negative impact on DNA recoverability with up to 98% of DNA lost over a 20-h sampling period. Pooling DNA from separate samples during extraction was proven to be feasible with margins of error below 30%.
ISSN: 1932-6203
DOI: 10.1371/journal.pone.0141158
Schools: School of Civil and Environmental Engineering 
Interdisciplinary Graduate School (IGS) 
Research Centres: Singapore Centre for Environmental Life Sciences Engineering 
Rights: © 2015 Luhung et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Fulltext Permission: open
Fulltext Availability: With Fulltext
Appears in Collections:CEE Journal Articles
SCELSE Journal Articles

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