4.1 Article

Comparing High-throughput Platforms for Sequencing the V4 Region of SSU-rDNA in Environmental Microbial Eukaryotic Diversity Surveys

期刊

JOURNAL OF EUKARYOTIC MICROBIOLOGY
卷 62, 期 3, 页码 338-345

出版社

WILEY
DOI: 10.1111/jeu.12187

关键词

454; abundances; Illumina; MiSeq; next-generation sequencing; pyrosequencing; Roche

资金

  1. Deutsche Forschungsgemeinschaft (DFG) [DU1319/1-1]

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High-throughput sequencing platforms are continuing to increase resulting read lengths, which is allowing for a deeper and more accurate depiction of environmental microbial diversity. With the nascent Reagent Kit v3, Illumina MiSeq now has the ability to sequence the eukaryotic hyper-variable V4 region of the SSU-rDNA locus with paired-end reads. Using DNA collected from soils with analyses of strictly- and nearly identical amplicons, here we ask how the new Illumina MiSeq data compares with what we can obtain with Roche/454 GS FLX with regard to quantity and quality, presence and absence, and abundance perspectives. We show that there is an easy qualitative transition from the Roche/454 to the Illumina MiSeq platforms. The ease of this transition is more nuanced quantitatively for low-abundant amplicons, although estimates of abundances are known to also vary within platforms.

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