Microvariation Artifacts Introduced by PCR and Cloning of Closely Related 16S rRNA Gene Sequences

A.G.C.L. Speksnijder, G.A. Kowalchuk, S. de Jong, E. Kline, J.R. Stephen, H.J. Laanbroek

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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
    Original languageEnglish
    Pages (from-to)469-472
    JournalApplied and Environmental Microbiology
    Volume67
    Issue number1
    DOIs
    Publication statusPublished - 2001

    Keywords

    • ribosomal-rna genes
    • polymerase chain-reaction
    • chimeric molecules
    • dna amplification
    • escherichia-coli
    • heteroduplexes
    • populations
    • coamplification
    • consequence
    • diversity

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    Speksnijder, A. G. C. L., Kowalchuk, G. A., de Jong, S., Kline, E., Stephen, J. R., & Laanbroek, H. J. (2001). Microvariation Artifacts Introduced by PCR and Cloning of Closely Related 16S rRNA Gene Sequences. Applied and Environmental Microbiology, 67(1), 469-472. https://doi.org/10.1128/AEM.67.1.469-472.2001