Microvariation artifacts introduced by PCR and cloning of closely related 16S rRNA gene sequences
Publication date
2001
Authors
Speksnijder, A.G.C.L.
Kowalchuk, G.A.
Jong, S. de
Kline, E.
Stephen, J.R.
Laanbroek, H.J.
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Article
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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.