Abstract：
:Next-generation sequencing has increased the coverage of microbial diversity surveys by orders of magnitude, but differentiating artifacts from rare environmental sequences remains a challenge. Clustering 16S rRNA sequences into operational taxonomic units (OTUs) organizes sequence data into groups of 97 % identity, helping to reduce data volumes and avoid analyzing sequencing artifacts by grouping them with real sequences. Here, we analyze sequence abundance distributions across environmental samples and show that 16S rRNA sequences of >99 % identity can represent functionally distinct microorganisms, rendering OTU clustering problematic when the goal is an accurate analysis of organism distribution. Strict postsequencing quality control (QC) filters eliminated the most prevalent artifacts without clustering. Further experiments proved that DNA polymerase errors in polymerase chain reaction (PCR) generate a significant number of substitution errors, most of which pass QC filters. Based on our findings, we recommend minimizing the number of PCR cycles in DNA library preparation and applying strict postsequencing QC filters to reduce the most prevalent artifacts while maintaining a high level of accuracy in diversity estimates. We further recommend correlating rare and abundant sequences across environmental samples, rather than clustering into OTUs, to identify remaining sequence artifacts without losing the resolution afforded by high-throughput sequencing.
journal_name
Microb Ecoljournal_title
Microbial ecologyauthors
Patin NV,Kunin V,Lidström U,Ashby MNdoi
10.1007/s00248-012-0145-4subject
Has Abstractpub_date
2013-04-01 00:00:00pages
709-19issue
3eissn
0095-3628issn
1432-184Xjournal_volume
65pub_type
杂志文章abstract：:TwoNostoc muscorum cyanophages were isolated from a domestic sewage in Kuwait. N-1L cyanophage had a hexagonal head with a long tail, while N-2S cyanophage was a short-tailed virus. N-1L cyanophage was active at 50°C and at acidic pH, compared with N-2S, which was more heat stable and active at pH 7.0. Seasonal variat...
journal_title：Microbial ecology
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更新日期：2011-11-01 00:00:00
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更新日期：2002-01-01 00:00:00
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更新日期：2021-01-01 00:00:00
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doi：10.1007/s00248-019-01340-8
更新日期：2019-05-01 00:00:00
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journal_title：Microbial ecology
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更新日期：2007-07-01 00:00:00
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journal_title：Microbial ecology
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更新日期：2000-08-01 00:00:00
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journal_title：Microbial ecology
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doi：10.1007/s00248-017-1062-3
更新日期：2018-04-01 00:00:00
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journal_title：Microbial ecology
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doi：10.1007/s00248-017-1126-4
更新日期：2018-08-01 00:00:00
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journal_title：Microbial ecology
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doi：10.1007/s00248-017-1137-1
更新日期：2018-08-01 00:00:00
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更新日期：1981-09-01 00:00:00
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更新日期：1988-07-01 00:00:00
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更新日期：2019-02-01 00:00:00
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更新日期：2015-11-01 00:00:00
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journal_title：Microbial ecology
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