Metabarcoding marine sediments: preparation of amplicon libraries
Fonseca, V.G.; Lallias, D. (2016). Metabarcoding marine sediments: preparation of amplicon libraries, in: Bourlat, S.J. (Ed.) Marine genomics: methods and protocols. Methods in Molecular Biology, 1452: pp. 183-196. https://dx.doi.org/10.1007/978-1-4939-3774-5_12
In: Bourlat, S.J. (Ed.) (2016). Marine genomics: methods and protocols. Methods in Molecular Biology, 1452. Humana Press/Springer Science+Business Media, Inc: New York. ISBN 978-1-4939-3772-1. 253 pp. https://dx.doi.org/10.1007/978-1-4939-3774-5, more
In: Methods in Molecular Biology. Humana Press. ISSN 1064-3745, more
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| Author keywords |
Metabarcoding; Marine eukaryotes; Environmental DNA; High-throughput sequencing; NGS; Molecular biodiversity; Illumina |
| Authors | | Top |
- Fonseca, V.G.
- Lallias, D.
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| Abstract |
The accurate assessment of community composition and ultimately species identification is of utmost importance in any ecological and evolutionary study. Advances in sequencing technologies have allowed the unraveling of levels of biodiversity never imagined before when applied to large-scale environmental DNA studies (also termed metabarcoding/metagenetics/metasystematics/environmental barcoding). Here, we describe a detailed protocol to assess eukaryotic biodiversity in marine sediments, identifying key steps that should not be neglected when preparing Next-Generation Sequencing (NGS) amplicon libraries: DNA extraction, multiple PCR amplification of DNA barcode markers with index/ tag-primers, and final Illumina MiSeq sequencing library preparation. |
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