KNAW Repository

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

Speksnijder, A.G.C.L. and Kowalchuk, G.A. and De Jong, S. and Kline, E. and Stephen, J.R. and Laanbroek, H.J. (2001) Microvariation artifacts introduced by PCR and cloning of closely related 16S rRNA gene sequences. Applied and Environmental Microbiology, 67, 469-472. ISSN 0099-2240.

[img]PDF - Published Version
Restricted to KNAW only

1507Kb

Official URL: http://dx.doi.org/10.1128/AEM.67.1.469-472.2001

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]

Item Type:Article
Institutes:Nederlands Instituut voor Ecologie (NIOO)
ID Code:11069
Deposited On:24 Nov 2011 01:00
Last Modified:31 Mar 2014 10:05

Repository Staff Only: item control page