Microvariation Artifacts Introduced by PCR and Cloning of Closely Related 16S rRNA Gene Sequences
Open Access
- 1 January 2001
- journal article
- Published by American Society for Microbiology in Applied and Environmental Microbiology
- Vol. 67 (1), 469-472
- https://doi.org/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.Keywords
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