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Preparation of Amplicon Libraries for Metabarcoding of Marine Eukaryotes Using Illumina MiSeq: The Dual-PCR Method

Chapter in book
Authors Sarah Bourlat
Quiterie Haenel
Jennie Finnman
Matthieu Leray
Published in Marine Genomics: Methods and Protocols
Pages 197-207
ISBN 978-1-4939-3772-1
Publisher Springer
Place of publication New York
Publication year 2016
Published at Department of marine sciences
Core Facilities, Genomics
Pages 197-207
Language en
Keywords Metabarcoding Biodiversity Microscopic eukaryotes Illumina MiSeq
Subject categories Biochemistry and Molecular Biology, Genetics

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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Utskriftsdatum: 2019-12-11