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Capturing diversity of marine heterotrophic protists: one cell at a time
Recent applications of culture-independent, molecular methods have revealed unexpectedly high diversity in a variety of functional and phylogenetic groups of microorganisms in the ocean. However, none of the existing research tools are free from significant limitations, such as PCR and cloning biase...
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Published in: | The ISME Journal 2011-04, Vol.5 (4), p.674-684 |
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description | Recent applications of culture-independent, molecular methods have revealed unexpectedly high diversity in a variety of functional and phylogenetic groups of microorganisms in the ocean. However, none of the existing research tools are free from significant limitations, such as PCR and cloning biases, low phylogenetic resolution and others. Here, we employed novel, single-cell sequencing techniques to assess the composition of small ( |
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Clone, but not SAG, libraries contained several large clusters of identical and nearly identical sequences of Dinophyceae, Cercozoa and Stramenopiles. Similar results were obtained using two alternative primer sets, suggesting that PCR biases may not be the only explanation for the observed patterns. Instead, differences in the number of 18S rRNA gene copies among the various protist taxa probably had a significant role in determining the PCR clone composition. These results show that single-cell sequencing has the potential to more accurately assess protistan community composition than previously established methods. 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Clone, but not SAG, libraries contained several large clusters of identical and nearly identical sequences of Dinophyceae, Cercozoa and Stramenopiles. Similar results were obtained using two alternative primer sets, suggesting that PCR biases may not be the only explanation for the observed patterns. Instead, differences in the number of 18S rRNA gene copies among the various protist taxa probably had a significant role in determining the PCR clone composition. These results show that single-cell sequencing has the potential to more accurately assess protistan community composition than previously established methods. 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subjects | 631/158/670 631/1647/514 631/326/2565 Aquatic Organisms - classification Aquatic Organisms - genetics Aquatic Organisms - isolation & purification Biodiversity Biomedical and Life Sciences Cloning Community composition Dinophyceae Ecology Environment Eukaryota - classification Eukaryota - genetics Eukaryota - isolation & purification Evolutionary Biology Flow Cytometry Gene Library Life Sciences Microbial Ecology Microbial Genetics and Genomics Microbiology Microorganisms Original original-article Phylogeny Polymerase Chain Reaction RNA, Ribosomal, 18S - genetics Seawater Sequence Analysis, DNA Stramenopiles |
title | Capturing diversity of marine heterotrophic protists: one cell at a time |
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