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Microvariation Artifacts Introduced by PCR and Cloning of Closely Related 16S rRNA Gene Sequences
Authors:Arjen G. C. L. Speksnijder   George A. Kowalchuk   Sander De Jong   Elizabeth Kline   John R. Stephen     Hendrikus J. Laanbroek
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.
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