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

• DOI: 10.1128/aem.67.1.469-472.2001
• Publication Date: 2002-07-27T10:00:58Z
• Authors: Arjen G. C. L. Sp...; George A. Kowalchuk; Sander De Jong; Elizabeth Kline; John R. Stephen; Hendrikus J. Laan...
• Keywords: clone (Java method); Cloning (programming); Library

ABSTRACT A defined template mixture of seven closely related 16S-rDNA clones was used in a PCR-cloning experiment to assess and track sources artifactual sequence variation 16S rDNA clone libraries. At least 14% the recovered contained aberrations. Artifact were polymerase errors, mutational hot spot, cloning heteroduplexes chimeras. These data may partially explain high degree microheterogeneity typical clusters detected environmental