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      Quality-filtering vastly improves diversity estimates from Illumina amplicon sequencing

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          Abstract

          High-throughput sequencing has revolutionized microbial ecology, but read quality remains a significant 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 from, and facilitating interpretation of Illumina sequencing results.

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          Author and article information

          Journal
          101215604
          32338
          Nat Methods
          Nat. Methods
          Nature methods
          1548-7091
          1548-7105
          21 November 2012
          02 December 2012
          January 2013
          01 July 2013
          : 10
          : 1
          : 57-59
          Affiliations
          [1 ]Department of Viticulture and Enology, University of California, Davis, CA, USA
          [2 ]Department of Food Science and Technology, University of California, Davis, CA, USA
          [3 ]Foods for Health Institute, University of California, Davis, CA, USA
          [4 ]Center for Genome Sciences and Systems Biology, Washington University School of Medicine, St. Louis, MO, USA
          [5 ]Microbial Systems & Communities, Genome Sequencing and Analysis Program, Broad Institute of MIT and Harvard, Cambridge, MA, USA
          [6 ]Department of Chemistry and Biochemistry, University of Colorado, Boulder, CO, USA
          [7 ]Howard Hughes Medical Institute, Boulder, CO, USA
          [8 ]Institute for Genomics and Systems Biology, Argonne National Laboratory, Argonne, IL, USA
          [9 ]Department of Computer Science, Northern Arizona University, Flagstaff, AZ, USA
          Author notes
          [* ]Corresponding author: Gregory Caporaso, Department of Computer Science, PO Box 15600, Northern Arizona University, Flagstaff, AZ, USA, (303) 523-5485, (303) 523-4015 (fax), gregcaporaso@ 123456gmail.com
          Article
          NIHMS420659
          10.1038/nmeth.2276
          3531572
          23202435
          b6a1255f-f6ed-4bbe-a4ad-c9451d27f49c

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          History
          Funding
          Funded by: National Human Genome Research Institute : NHGRI
          Award ID: U54 HG004969 || HG
          Funded by: National Institute of Child Health & Human Development : NICHD
          Award ID: R01 HD059127 || HD
          Funded by: National Institute of Diabetes and Digestive and Kidney Diseases : NIDDK
          Award ID: P01 DK078669 || DK
          Funded by: Howard Hughes Medical Institute :
          Award ID: || HHMI_
          Categories
          Article

          Life sciences
          Life sciences

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