title: Preparation of Amplicon Libraries for Metabarcoding of Marine Eukaryotes Using Illumina MiSeq: The Adapter Ligation Method.
abstract: Amplicon-based studies of marine microscopic eukaryotes, also referred to as metabarcoding studies, can be performed to analyze patterns of biodiversity or predator-prey interactions targeting the mitochondrial cytochrome oxidase 1 (CO1) or the small ribosomal subunit (SSU) markers. Because high-throughput sequencing (HTS) Illumina platforms provide millions of reads per run, hundreds of samples may be sequenced simultaneously. This protocol details the preparation of multiplexed amplicon libraries for Illumina MiSeq sequencing. We describe a strategy for sample multiplexing using a combination of tailed PCR primers and ligation of indexed adapters.
authors.affiliation: Department of Invertebrate Zoology, National Museum of Natural History, Smithsonian Institution, 10th and Constitution Ave., NW, Washington, DC, 20560, USA. leray.upmc@gmail.com.
authors.forename: Matthieu
authors.initials: M
authors.lastname: Leray
authors.affiliation: Department of Biological and Environmental Sciences, University of Gothenburg, Box 461, Gothenburg, SE-413 19, Sweden.
authors.forename: Quiterie
authors.initials: Q
authors.lastname: Haenel
authors.affiliation: Department of Marine Sciences, University of Gothenburg, Gothenburg, Sweden.
authors.forename: Sarah J
authors.initials: SJ
authors.lastname: Bourlat
mesh_terms: D003576:Electron Transport Complex IV / Q000235:genetics; D056890:Eukaryota / Q000235:genetics; D059014:High-Throughput Nucleotide Sequencing; D008386:Marine Biology / Q000379:methods*; D054679:Ribosome Subunits, Small / Q000235:genetics; D012984:Software; D014871:Water Microbiology
chemical_list: D003576:Electron Transport Complex IV
keywords: Eukaryotes; Illumina MiSeq; Metabarcoding; Multiplexing; Tags; TruSeq