A Method for Studying Protistan Diversity Using Massively Parallel Sequencing of V9 Hypervariable Regions of Small-Subunit Ribosomal RNA Genes

BACKGROUND: Massively parallel pyrosequencing of amplicons from the V6 hypervariable regions of small-subunit (SSU) ribosomal RNA (rRNA) genes is commonly used to assess diversity and richness in bacterial and archaeal populations. Recent advances in pyrosequencing technology provide read lengths of up to 240 nucleotides. Amplicon pyrosequencing can now be applied to longer variable regions of the SSU rRNA gene including the V9 region in eukaryotes. METHODOLOGY/PRINCIPAL FINDINGS: We present a protocol for the amplicon pyrosequencing of V9 regions for eukaryotic environmental samples for biodiversity inventories and species richness estimation. The International Census of Marine Microbes (ICoMM) and the…

• NS
  National Science Foundation

  Awards: NSF/OCE 0430724, 0717390, 0217282, P50 ES012742, 0430724, OPP 0217282
• WM
  W. M. Keck Foundation
• NI
  National Institutes of Health
• NI
  National Institute of Environmental Health Sciences

  Awards: P50 ES012742, 1 P50 ES012742
• DO
  Division of Environmental Biology