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Methods Mol Biol. 2016;1452:197-207. doi: 10.1007/978-1-4939-3774-5_13.

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

Author information

1
Department of Marine Sciences, University of Gothenburg, Box 463, SE-405 30, Gothenburg, Sweden. sarah.bourlat@marine.gu.se.
2
Department of Biological and Environmental Sciences, University of Gothenburg, Box 461, SE-413 19, Gothenburg, Sweden.
3
Sahlgrenska Academy, Gothenburg University Genomics Core Facility, Gothenburg, Sweden.
4
National Museum of Natural History, Smithsonian Institution, Washington, DC, USA.

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.

KEYWORDS:

Biodiversity; Illumina MiSeq; Metabarcoding; Microscopic eukaryotes

PMID:
27460379
DOI:
10.1007/978-1-4939-3774-5_13
[Indexed for MEDLINE]

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