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

J Eukaryot Microbiol. 2015 May-Jun;62(3):338-45. doi: 10.1111/jeu.12187. Epub 2014 Nov 20.

Abstract

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.

Keywords: 454; Illumina; MiSeq; Roche; abundances; next-generation sequencing; pyrosequencing.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biota*
  • Environmental Microbiology*
  • High-Throughput Nucleotide Sequencing / methods*
  • RNA, Ribosomal, 18S / genetics*

Substances

  • RNA, Ribosomal, 18S