4.8 Article

Quality-filtering vastly improves diversity estimates from Illumina amplicon sequencing

Journal

NATURE METHODS
Volume 10, Issue 1, Pages 57-U11

Publisher

NATURE PUBLISHING GROUP
DOI: 10.1038/NMETH.2276

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Funding

  1. US National Institutes of Health [NIH DK78669, NIH R01HD059127, NIH U54HG004969]
  2. Juvenile Diabetes Research Fund
  3. Crohn's and Colitis Foundation of America
  4. Howard Hughes Medical Institute
  5. Dannon Probiotics Fellow Program (The Dannon Company)
  6. Wine Spectator scholarship

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High-throughput sequencing has revolutionized microbial ecology, but read quality remains a considerable barrier to accurate taxonomy assignment and alpha-diversity assessment for microbial communities. We demonstrate that high-quality read length and abundance are the primary factors differentiating correct from erroneous reads produced by Illumina GAIIx, HiSeq and MiSeq instruments. We present guidelines for user-defined quality-filtering strategies, enabling efficient extraction of high-quality data and facilitating interpretation of Illumina sequencing results.

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