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

被引：182
作者：
Speksnijder, AGCL
Kowalchuk, GA
De Jong, S
Kline, E
Stephen, JR
Laanbroek, HJ
机构：
[1] Nat Hist Museum, Dept Zool, London SW7 5BD, England
[2] Netherlands Inst Ecol, Ctr Terr Ecol, Dept Plant Microorganism Interact, NL-6666 ZG Heteren, Netherlands
[3] Netherlands Inst Ecol, Ctr Limnol, Dept Microbial Ecol, NL-3136 AC Nieuwersluis, Netherlands
[4] Univ Tennessee, Ctr Environm Biotechnol, Knoxville, TN 37932 USA
来源：
APPLIED AND ENVIRONMENTAL MICROBIOLOGY | 2001年 / 67卷 / 01期
基金：
美国国家科学基金会;
关键词：
D O I：
10.1128/AEM.67.1.469-472.2001
中图分类号：
Q81 [生物工程学（生物技术）]; Q93 [微生物学];
学科分类号：
071005 ; 0836 ; 090102 ; 100705 ;
摘要：
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.
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收藏
页码：469 / 472
页数：4
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