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

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.

SUBMITTER: Speksnijder AG 

PROVIDER: S-EPMC92603 | biostudies-literature | 2001 Jan

REPOSITORIES: biostudies-literature