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      DNA extraction for streamlined metagenomics of diverse environmental samples.

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          Abstract

          A major bottleneck for metagenomic sequencing is rapid and efficient DNA extraction. Here, we compare the extraction efficiencies of three magnetic bead-based platforms (KingFisher, epMotion, and Tecan) to a standardized column-based extraction platform across a variety of sample types, including feces, oral, skin, soil, and water. Replicate sample plates were extracted and prepared for 16S rRNA gene amplicon sequencing in parallel to assess extraction bias and DNA quality. The data demonstrate that any effect of extraction method on sequencing results was small compared with the variability across samples; however, the KingFisher platform produced the largest number of high-quality reads in the shortest amount of time. Based on these results, we have identified an extraction pipeline that dramatically reduces sample processing time without sacrificing bacterial taxonomic or abundance information.

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          Author and article information

          Journal
          BioTechniques
          BioTechniques
          Informa UK (Informa Life Sciences)
          1940-9818
          0736-6205
          Jun 01 2017
          : 62
          : 6
          Affiliations
          [1 ] Department of Pediatrics, University of California, San Diego, La Jolla, CA.
          Article
          000114559
          10.2144/000114559
          28625159
          a4181d68-5317-483b-a588-326b015fe80a
          History

          metagenomics,high throughput,DNA extraction,microbiom
          metagenomics, high throughput, DNA extraction, microbiom

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