Preparation of amplicon libraries for metabarcoding of marine eukaryotes using Illumina MiSeq: the dual- PCR method
Erscheinungsjahr:
2016
Vollständiger Titel:
Preparation of amplicon libraries for metabarcoding of marine eukaryotes using Illumina MiSeq: the dual- PCR method
ZFMK-Autorinnen / ZFMK-Autoren:
Org. Einordnung:
Publiziert in:
Methods in Molecular Biology
Publikationstyp:
Beitrag zu Sammelwerk
DOI Name:
10.1007/978-1-4939-3774-5_13
Keywords:
Metabarcoding; Biodiversity; Microscopic eukaryotes; Illumina MiSeq
Bibliographische Angaben:
Bourlat, S. J., Haenel, Q., Finnman, J., Leray, M. (2016): Preparation of amplicon libraries for metabarcoding of marine eukaryotes using Illumina MiSeq: the dual- PCR method. In: Bourlat, S. J. (ed.): Marine Genomics - Methods and protocols. - Springer (New York). pp. 197-208. https://doi.org/10.1007/978-1-4939-3774-5_13
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.
