Microvariation artifacts introduced by PCR and cloning of closely related 16S rRNA gene sequences

Arjen Speksnijder, G.A. Kowalchuk, S. De Jong, E. Kline, J.R. Stephen, H.J. Laanbroek

    Research output: Contribution to journal/periodicalArticleScientificpeer-review

    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: Ribosomal-rna genes; polymerase chain-reaction; chimeric molecules; dna amplification; escherichia-coli heteroduplexes; populations; coamplification; consequence; diversity]
    Original languageEnglish
    Pages (from-to)469-472
    JournalApplied and Environmental Microbiology
    Volume67
    Issue number1
    DOIs
    Publication statusPublished - 2001

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