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.

引用

页码：469 / 472

页数：4

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