Microvariation Artifacts Introduced by PCR and Cloning of Closely Related 16S rRNA Gene Sequences-Reference-Cited by-同舟云学术

Microvariation Artifacts Introduced by PCR and Cloning of Closely Related 16S rRNA Gene Sequences

Published:2001-01 Issue:1 Volume:67 Page:469-472
ISSN:0099-2240
Container-title:Applied and Environmental Microbiology
language:en
Short-container-title:Appl Environ Microbiol

Author:

Speksnijder Arjen G. C. L.¹²,Kowalchuk George A.³,De Jong Sander²,Kline Elizabeth⁴,Stephen John R.⁴,Laanbroek Hendrikus J.²

Affiliation:

1. Department of Zoology, Natural History Museum, South Kensington, London SW7 5BD, United Kingdom1;

2. Department of Microbial Ecology, Centre for Limnology, Netherlands Institute of Ecology, 3136 AC Nieuwersluis,2 The Netherlands; and

3. Department of Plant-Microorganism Interactions, Centre for Terrestrial Ecology, Netherlands Institute of Ecology, 6666 ZG Heteren,3 and

4. Center for Environmental Biotechnology, University of Tennessee, Knoxville, Tennessee 37932-25754

Abstract

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.

Publisher

American Society for Microbiology

Subject

Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology

Link

https://journals.asm.org/doi/pdf/10.1128/AEM.67.1.469-472.2001

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