Abstract
Methods to obtain high-quality assembled genomic information of rare and unclassified member species in complex microbial communities remain a high priority in microbial ecology. Additionally, the supplementation of three-dimensional spatial information that highlights the morphology and spatial interaction would provide additional insights to its ecological role in the community. Fluorescent in-situ hybridization (FISH) coupling with fluorescence-activated cell sorting (FACS) is a powerful tool that enables the detection, visualization, and separation of low-abundance microbial members in samples containing complex microbial compositions. Here, we have described the workflow from designing the appropriate FISH probes from metagenomics or metatranscriptomics datasets to the preparation and treatment of samples to be used in FISH-FACS procedures.
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Acknowledgments
This research was supported by the Singapore National Research Foundation (NRF) and the Ministry of Education (MOE) under the Research Centre of Excellence Programme. Dr. Shi Ming Tan was supported by the Singapore NRF Environmental and Water Technologies (EWT) PhD Scholarship. We thank Larry Liew for performing the sampling of activated sludge and the technical staff from the SCELSE sequencing laboratory for their assistance.
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The authors declare no competing financial or nonfinancial interests.
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Yung, P.Y.M., Tan, S.M. (2023). Targeted Enrichment of Low-Abundance and Uncharacterized Taxon Members in Complex Microbial Community with Primer-Free FISH Probes Designed from Next Generation Sequencing Dataset. In: Mitra, S. (eds) Metagenomic Data Analysis. Methods in Molecular Biology, vol 2649. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3072-3_16
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DOI: https://doi.org/10.1007/978-1-0716-3072-3_16
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