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Novel primer sets for next generation sequencing-based analyses of water quality
Journal article   Open access   Peer reviewed

Novel primer sets for next generation sequencing-based analyses of water quality

E. Lee, M.S. Khurana, A.S. Whiteley, P.T. Monis, A. Bath, C. Gordon, U.M. Ryan and A. Paparini
PLoS ONE, Vol.12(1), e0170008
2017
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Abstract

Next generation sequencing (NGS) has rapidly become an invaluable tool for the detection, identification and relative quantification of environmental microorganisms. Here, we demonstrate two new 16S rDNA primer sets, which are compatible with NGS approaches and are primarily for use in water quality studies. Compared to 16S rRNA gene based universal primers, in silico and experimental analyses demonstrated that the new primers showed increased specificity for the Cyanobacteria and Proteobacteria phyla, allowing increased sensitivity for the detection, identification and relative quantification of toxic bloom-forming microalgae, microbial water quality bioindicators and common pathogens. Significantly, Cyanobacterial and Proteobacterial sequences accounted for ca. 95% of all sequences obtained within NGS runs (when compared to ca. 50% with standard universal NGS primers), providing higher sensitivity and greater phylogenetic resolution of key water quality microbial groups. The increased selectivity of the new primers allow the parallel sequencing of more samples through reduced sequence retrieval levels required to detect target groups, potentially reducing NGS costs by 50% but still guaranteeing optimal coverage and species discrimination.

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UN Sustainable Development Goals (SDGs)

This output has contributed to the advancement of the following goals:

#6 Clean Water and Sanitation

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Collaboration types
Domestic collaboration
Citation topics
3 Agriculture, Environment & Ecology
3.2 Marine Biology
3.2.216 Lake Ecosystems
Web Of Science research areas
Microbiology
ESI research areas
Biology & Biochemistry
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