title: Preparation of Amplicon Libraries for Metabarcoding of Marine Eukaryotes Using Illumina MiSeq: The Dual-PCR Method.
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.
authors.affiliation: Department of Marine Sciences, University of Gothenburg, Box 463, SE-405 30, Gothenburg, Sweden. sarah.bourlat@marine.gu.se.
authors.forename: Sarah J
authors.initials: SJ
authors.lastname: Bourlat
authors.affiliation: Department of Biological and Environmental Sciences, University of Gothenburg, Box 461, SE-413 19, Gothenburg, Sweden.
authors.forename: Quiterie
authors.initials: Q
authors.lastname: Haenel
authors.affiliation: Sahlgrenska Academy, Gothenburg University Genomics Core Facility, Gothenburg, Sweden.
authors.forename: Jennie
authors.initials: J
authors.lastname: Finnman
authors.affiliation: National Museum of Natural History, Smithsonian Institution, Washington, DC, USA.
authors.forename: Matthieu
authors.initials: M
authors.lastname: Leray
mesh_terms: D044822:Biodiversity; D058893:DNA Barcoding, Taxonomic / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA
keywords: Biodiversity; Illumina MiSeq; Metabarcoding; Microscopic eukaryotes