Microvariation artifacts introduced by PCR and cloning of closely related 16S rRNA gene sequences.
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.
Speksnijder
A G
AG
Department of Zoology, Natural History Museum, South Kensington, London SW7 5BD, United Kingdom. a.speksnijder@abdn.ac.uk
Kowalchuk
G A
GA
De Jong
S
S
Kline
E
E
Stephen
J R
JR
Laanbroek
H J
HJ
D016477:Artifacts; D001419:Bacteria / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods*; D015723:Gene Library; D020459:Genes, rRNA; D014644:Genetic Variation; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D017422:Sequence Analysis, DNA
D012336:RNA, Ribosomal, 16S