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. - \ 2001
Applied and Environmental Microbiology 67 (2001)1. - ISSN 0099-2240 - p. 469 - 472.
ribosomal-rna genes - polymerase chain-reaction - chimeric molecules - dna amplification - escherichia-coli - heteroduplexes - populations - coamplification - consequence - diversity
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