Practical innovations for high-throughput amplicon sequencing

Nat Methods. 2013 Oct;10(10):999-1002. doi: 10.1038/nmeth.2634. Epub 2013 Sep 1.


We describe improvements for sequencing 16S ribosomal RNA (rRNA) amplicons, a cornerstone technique in metagenomics. Through unique tagging of template molecules before PCR, amplicon sequences can be mapped to their original templates to correct amplification bias and sequencing error with software we provide. PCR clamps block amplification of contaminating sequences from a eukaryotic host, thereby substantially enriching microbial sequences without introducing bias.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteria* / classification
  • Bacteria* / genetics
  • Classification / methods*
  • High-Throughput Nucleotide Sequencing / methods*
  • Metagenome*
  • RNA, Ribosomal, 16S / genetics*
  • Software
  • Soil Microbiology*


  • RNA, Ribosomal, 16S