Microvariation artifacts introduced by PCR and cloning of closely related 16S rRNA gene sequences

Appl Environ Microbiol. 2001 Jan;67(1):469-72. doi: 10.1128/AEM.67.1.469-472.2001.

Abstract

A defined template mixture of seven closely related 16S-rDNA clones was used in a PCR-cloning experiment to assess and track sources of artifactual sequence variation in 16S rDNA clone libraries. At least 14% of the recovered clones contained aberrations. Artifact sources were polymerase errors, a mutational hot spot, and cloning of heteroduplexes and chimeras. These data may partially explain the high degree of microheterogeneity typical of sequence clusters detected in environmental clone libraries.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Artifacts*
  • Bacteria / genetics*
  • Base Sequence
  • Cloning, Molecular / methods*
  • Gene Library
  • Genes, rRNA
  • Genetic Variation*
  • Molecular Sequence Data
  • Polymerase Chain Reaction / methods*
  • RNA, Ribosomal, 16S / genetics*
  • Sequence Analysis, DNA

Substances

  • RNA, Ribosomal, 16S