Microvariation artifacts introduced by PCR and cloning of closely related 16S rRNA gene sequences
- Speksnijder, A.G.C.L.
- Kowalchuk, G.A.
- Jong, S. de
- Kline, E.
- Stephen, J.R.
- Laanbroek, H.J.
Publication date
January 2001
DOI Publisher
American Society for MicrobiologyAbstract
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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