The DOI matches its Crossref record; 29 of the 29 references it cites were found in a registry or an index.
Frequency of formation of chimeric molecules as a consequence of PCR coamplification of 16S rRNA genes from mixed bacterial genomes
Open accessAbstract
PCR is routinely used in amplification and cloning of rRNA genes from environmental DNA samples for studies of microbial community structure and identification of novel organisms. There have been concerns about generation of chimeric sequences as a consequence of PCR coamplification of highly conserved genes, because such sequences may lead to reports of nonexistent organisms. To quantify the frequency of chimeric molecule formation, mixed genomic DNAs from eight actinomycete species whose 16S rRNA sequences had been determined were used for PCR coamplification of 16S rRNA genes. A large number of cloned 16S ribosomal DNAs were examined by sequence analysis, and chimeric molecules were identified by multiple-sequence alignment with reference species. Here, we report that the level of occurrence of chimeric sequences after 30 cycles of PCR amplification was 32%. We also show that PCR-induced chimeras were formed between different rRNA gene copies from the same organism. Because of the wide use of PCR for direct isolation of 16S rRNA sequences from environmental DNA to assess microbial diversity, the extent of chimeric molecule formation deserves serious attention.
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Cites
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Cited by
UCHIME improves sensitivity and speed of chimera detection
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Culture-Independent Analysis of Gut Bacteria: the Pig Gastrointestinal Tract Microbiota Revisited
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Metadata
- Authors
- Wang, G. C., & Wang, Y.
- Venue
- Applied and Environmental Microbiology
- Publisher
- American Society for Microbiology
- Published
- 1997-12
- Volume
- 63
- Issue
- 12
- Pages
- 4645-4650
- ISSN
- 0099-2240 (ISSN-L)
- 1098-5336
- OpenAlex
- W2106944866
- Cited by
- 337
- References
- 29
- FWCI
- 12.16
- Type
- journal article
- Language
- en
- Open access
- green
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- Wang, G. C., & Wang, Y. (1997). Frequency of formation of chimeric molecules as a consequence of PCR coamplification of 16S rRNA genes from mixed bacterial genomes. Applied and Environmental Microbiology. https://doi.org/10.1128/aem.63.12.4645-4650.1997
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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
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