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Preparation of amplicon libraries for metabarcoding of marine eukaryotes using Illumina MiSeq: the adaptor ligation method

AutorInnen: 
Leray, M., Haenel, Q., Bourlat, S.J.
Erscheinungsjahr: 
2016
Vollständiger Titel: 
Preparation of amplicon libraries for metabarcoding of marine eukaryotes using Illumina MiSeq: the adaptor ligation method
Autor/-innen des ZFMK: 
Publiziert in: 
Methods in Molecular Biology
Publikationstyp: 
Beitrag zu Sammelwerk
DOI Name: 
10.1007/978-1-4939-3774-5_14
Keywords: 
Metabarcoding; Eukaryotes: Illumina MiSeq; Multiplexing; Tags; TruSeq
Bibliographische Angaben: 
Leray, M., Haenel, Q., Bourlat, S.J. (2016): Preparation of amplicon libraries for metabarcoding of marine eukaryotes using Illumina MiSeq: the adaptor ligation method. In: Bourlat, S. J. (ed.): Marine Genomics - Methods and protocols. - Springer (New York). pp. 209-218. https://doi.org/10.1007/978-1-4939-3774-5_14
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.