Cited by 100Microbial Community Ecology and PhysiologyGenomics and Phylogenetic StudiesEnvironmental DNA in Biodiversity Studies

Advantage of Using Inosine at the 3′ Termini of 16S rRNA Gene Universal Primers for the Study of Microbial Diversity

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Ben-Dov, Eitan, Shapiro, Orr H., Siboni, Nachshon, Kushmaro, Ariel·2006·Applied and Environmental Microbiology

Abstract

To overcome the shortcomings of universal 16S rRNA gene primers 8F and 907R when studying the diversity of complex microbial communities, the 3′ termini of both primers were replaced with inosine. A comparison of the clone libraries derived using both primer sets showed seven bacterial phyla amplified by the altered primer set (8F-I/907R-I) whereas the original set amplified sequences belonging almost exclusively to Proteobacteria(95.8%). Sequences belonging to Firmicutes(42.6%) and Thermotogae(9.3%) were more abundant in a library obtained by using 8F-I/907R-I at a PCR annealing temperature of 54°C, while Proteobacteria sequences were more frequent (62.7%) in a library obtained at 50°C, somewhat resembling the result obtained using the original primer set. The increased diversity revealed by using primers 8F-I/907R-I confirms the usefulness of primers with inosine at the 3′ termini in studying the microbial diversity of environmental samples.

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Crossref · 10.1128/aem.00849-06

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Authors
Ben-Dov, E., Shapiro, O. H., Siboni, N., & Kushmaro, A.
Affiliations
  • Department of Biotechnology Engineering
  • Ben-Gurion University of the Negev (IL)
Venue
Applied and Environmental Microbiology
Published
2006-11
Volume
72
Issue
11
Pages
6902-6906
ISSN
  • 0099-2240 (ISSN-L)
  • 1098-5336
OpenAlex
W2107352655
Cited by
100
References
31
Citations by year
2026: 1 · 2025: 2 · 2024: 3 · 2023: 4 · 2022: 3 · 2021: 6 · 2020: 5 · 2019: 2 · 2018: 3 · 2017: 4 · 2016: 3 · 2015: 6 · 2014: 4 · 2013: 7 · 2012: 8
FWCI
7.23
Type
journal article
Language
en
Open access
Locations
  • Applied and Environmental Microbiology (publishedVersion)
  • Europe PMC (PubMed Central) (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
  • UTS ePRESS (University of Technology Sydney) (submittedVersion)
Topics
  • Physical Sciences › Environmental Science › Ecology › Microbial Community Ecology and Physiology
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Phylogenetic Studies
  • Physical Sciences › Environmental Science › Ecology › Environmental DNA in Biodiversity Studies
Keywords
16S rRNA gene, universal primers, microbial diversity, bacterial phyla, Proteobacteria, Firmicutes, Thermotogae, PCR annealing temperature, clone libraries, environmental microbial communities
MeSH terms
Bacteria, Industrial Waste, Inosine, Molecular Sequence Data, Phylogeny, RNA, Ribosomal, 16S, Sodium Chloride, Genetic Variation, Water Microbiology, Polymerase Chain Reaction, Sequence Analysis, DNA, Ecosystem, DNA Primers, Genes, rRNA

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