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      Barcoded primers used in multiplex amplicon pyrosequencing bias amplification.

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          Abstract

          "Barcode-tagged" PCR primers used for multiplex amplicon sequencing generate a thus-far-overlooked amplification bias that produces variable terminal restriction fragment length polymorphism (T-RFLP) and pyrosequencing data from the same environmental DNA template. We propose a simple two-step PCR approach that increases reproducibility and consistently recovers higher genetic diversity in pyrosequencing libraries.

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          Author and article information

          Journal
          Appl Environ Microbiol
          Applied and environmental microbiology
          American Society for Microbiology
          1098-5336
          0099-2240
          Nov 2011
          : 77
          : 21
          Affiliations
          [1 ] Department of Microbial Ecology, Vienna Ecology Center, Faculty of Life Sciences, University of Vienna, Althanstrasse 14, A-1090 Vienna, Austria.
          Article
          AEM.05220-11
          10.1128/AEM.05220-11
          3209180
          21890669
          cb2d6158-9418-49a7-896a-b2df52e33730
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