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      Continuous base identification for single-molecule nanopore DNA sequencing.

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          Abstract

          A single-molecule method for sequencing DNA that does not require fluorescent labelling could reduce costs and increase sequencing speeds. An exonuclease enzyme might be used to cleave individual nucleotide molecules from the DNA, and when coupled to an appropriate detection system, these nucleotides could be identified in the correct order. Here, we show that a protein nanopore with a covalently attached adapter molecule can continuously identify unlabelled nucleoside 5'-monophosphate molecules with accuracies averaging 99.8%. Methylated cytosine can also be distinguished from the four standard DNA bases: guanine, adenine, thymine and cytosine. The operating conditions are compatible with the exonuclease, and the kinetic data show that the nucleotides have a high probability of translocation through the nanopore and, therefore, of not being registered twice. This highly accurate tool is suitable for integration into a system for sequencing nucleic acids and for analysing epigenetic modifications.

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          Author and article information

          Journal
          Nat Nanotechnol
          Nature nanotechnology
          Springer Science and Business Media LLC
          1748-3395
          1748-3387
          Apr 2009
          : 4
          : 4
          Affiliations
          [1 ] Oxford Nanopore Technologies Ltd, Begbroke Science Park, Sandy Lane, Oxford OX5 1PF, UK.
          Article
          nnano.2009.12
          10.1038/nnano.2009.12
          19350039
          6ac18058-c19e-4c89-9eaf-c4565289345d

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