Abstract
This protocol details the preparation of multiplexed amplicon libraries for metabarcoding (amplicon-based) studies of microscopic marine eukaryotes. Metabarcoding studies, based on the amplification of a taxonomically informative marker from a collection of organisms or an environmental sample, can be performed to analyze biodiversity patterns or predator–prey interactions. For Metazoa, we use the mitochondrial cytochrome oxidase 1 (CO1) or the small ribosomal subunit (SSU) markers. Here, we describe a strategy for the preparation of multiplexed Illumina MiSeq libraries using a dual-PCR approach for the addition of index and adaptor sequences.
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Acknowledgements
This work was supported by Swedish research council grant C0344601 to S.J.B. Financial support to M.L. was provided by the Sant Chair and the Smithsonian Tennenbaum Marine Observatories Network, for which this Contribution No. 4. We would like to thank the Genomics Core Facility platform, at the Sahlgrenska Academy, University of Gothenburg.
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Bourlat, S.J., Haenel, Q., Finnman, J., Leray, M. (2016). Preparation of Amplicon Libraries for Metabarcoding of Marine Eukaryotes Using Illumina MiSeq: The Dual-PCR Method. In: Bourlat, S. (eds) Marine Genomics. Methods in Molecular Biology, vol 1452. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3774-5_13
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DOI: https://doi.org/10.1007/978-1-4939-3774-5_13
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Publisher Name: Humana Press, New York, NY
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