Microvariation Artifacts Introduced by PCR and Cloning of Closely Related 16S rRNA Gene Sequences
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| Published in | Applied and Environmental Microbiology Vol. 67; no. 1; pp. 469 - 472 |
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| Main Authors | , , , , , |
| Format | Journal Article |
| Language | English |
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Washington, DC
American Society for Microbiology
01.01.2001
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| AbstractList | 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. 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. Classifications Services AEM Citing Articles Google Scholar PubMed Related Content Social Bookmarking CiteULike Delicious Digg Facebook Google+ Mendeley Reddit StumbleUpon Twitter current issue Spotlights in the Current Issue AEM About AEM Subscribers Authors Reviewers Advertisers Inquiries from the Press Permissions & Commercial Reprints ASM Journals Public Access Policy AEM RSS Feeds 1752 N Street N.W. • Washington DC 20036 202.737.3600 • 202.942.9355 fax • journals@asmusa.org Print ISSN: 0099-2240 Online ISSN: 1098-5336 Copyright © 2014 by the American Society for Microbiology. For an alternate route to AEM .asm.org, visit: AEM 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. 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 |
| Author | Elizabeth Kline Hendrikus J. Laanbroek Arjen G. C. L. Speksnijder John R. Stephen Sander De Jong George A. Kowalchuk |
| AuthorAffiliation | Department of Zoology, Natural History Museum, South Kensington, London SW7 5BD, United Kingdom 1 ; Department of Plant-Microorganism Interactions, Centre for Terrestrial Ecology, Netherlands Institute of Ecology, 6666 ZG Heteren, 3 and Department of Microbial Ecology, Centre for Limnology, Netherlands Institute of Ecology, 3136 AC Nieuwersluis, 2 The Netherlands; and Center for Environmental Biotechnology, University of Tennessee, Knoxville, Tennessee 37932-2575 4 |
| AuthorAffiliation_xml | – name: Department of Zoology, Natural History Museum, South Kensington, London SW7 5BD, United Kingdom 1 ; Department of Plant-Microorganism Interactions, Centre for Terrestrial Ecology, Netherlands Institute of Ecology, 6666 ZG Heteren, 3 and Department of Microbial Ecology, Centre for Limnology, Netherlands Institute of Ecology, 3136 AC Nieuwersluis, 2 The Netherlands; and Center for Environmental Biotechnology, University of Tennessee, Knoxville, Tennessee 37932-2575 4 |
| Author_xml | – sequence: 1 givenname: Arjen G. C. L surname: SPEKSNIJDER fullname: SPEKSNIJDER, Arjen G. C. L organization: Department of Zoology, Natural History Museum, South Kensington, London SW7 5BD, United Kingdom – sequence: 2 givenname: George A surname: KOWALCHUK fullname: KOWALCHUK, George A organization: Department of Microbial Ecology, Centre for Limnology, Netherlands Institute of Ecology, 3136 AC, Nieuwersluis, Netherlands – sequence: 3 givenname: Sander surname: DE JONG fullname: DE JONG, Sander organization: Department of Plant-Microorganism Interactions, Centre for Terrestrial Ecology, Netherlands Institute of Ecology, 6666 ZG Heteren, Netherlands – sequence: 4 givenname: Elizabeth surname: KLINE fullname: KLINE, Elizabeth organization: Center for Environmental Biotechnology, University of Tennessee, Knoxville, Tennessee 37932-2575, United States – sequence: 5 givenname: John R surname: STEPHEN fullname: STEPHEN, John R organization: Center for Environmental Biotechnology, University of Tennessee, Knoxville, Tennessee 37932-2575, United States – sequence: 6 givenname: Hendrikus J surname: LAANBROEK fullname: LAANBROEK, Hendrikus J organization: Department of Plant-Microorganism Interactions, Centre for Terrestrial Ecology, Netherlands Institute of Ecology, 6666 ZG Heteren, Netherlands |
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Reddit... 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... 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... |
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| StartPage | 469 |
| SubjectTerms | Bacteria - genetics Bacteriological methods and techniques used in bacteriology Bacteriology Base Sequence Biological and medical sciences chimeric molecules Cloning Cloning, Molecular - methods coamplification consequence Deoxyribonucleic acid diversity DNA dna amplification Environment escherichia-coli Fundamental and applied biological sciences. Psychology Gene Library Genes Genes, rRNA Genetic Variation heteroduplexes Microbial Ecology Microbiology Molecular Sequence Data Polymerase Chain Reaction - methods polymerase chain-reaction populations ribosomal-rna genes RNA, Ribosomal, 16S - genetics rRNA 16S Sequence Analysis, DNA |
| Title | Microvariation Artifacts Introduced by PCR and Cloning of Closely Related 16S rRNA Gene Sequences |
| URI | http://aem.asm.org/content/67/1/469.abstract https://www.ncbi.nlm.nih.gov/pubmed/11133483 https://www.proquest.com/docview/205966270 https://search.proquest.com/docview/17829404 https://search.proquest.com/docview/70548847 https://pubmed.ncbi.nlm.nih.gov/PMC92603 http://www.narcis.nl/publication/RecordID/oai:library.wur.nl:wurpubs%2F347840 |
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