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      Real-time selective sequencing using nanopore technology

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      1 , 2 , 1 , 1
      Nature methods

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          Abstract

          The Oxford Nanopore MinION sequences DNA by sensing changes in electrical current flow in real-time as molecules traverse nanopores. Optionally, the voltage across specific nanopores can be reversed, ejecting the DNA molecule. This enables “Read Until”, the selection of specific DNA molecules for sequencing. We use dynamic time warping to match reads to reference, selecting regions of small genomes, individual amplicons, or normalization of the amplicon set. This first demonstration of direct selection of specific DNA molecules in real-time enables many novel future applications.

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          Identification of common molecular subsequences.

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            Binary codes capable of correcting deletions, insertions, and reversals

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              MinION nanopore sequencing identifies the position and structure of a bacterial antibiotic resistance island.

              Short-read, high-throughput sequencing technology cannot identify the chromosomal position of repetitive insertion sequences that typically flank horizontally acquired genes such as bacterial virulence genes and antibiotic resistance genes. The MinION nanopore sequencer can produce long sequencing reads on a device similar in size to a USB memory stick. Here we apply a MinION sequencer to resolve the structure and chromosomal insertion site of a composite antibiotic resistance island in Salmonella Typhi Haplotype 58. Nanopore sequencing data from a single 18-h run was used to create a scaffold for an assembly generated from short-read Illumina data. Our results demonstrate the potential of the MinION device in clinical laboratories to fully characterize the epidemic spread of bacterial pathogens.
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                Author and article information

                Journal
                101215604
                32338
                Nat Methods
                Nat. Methods
                Nature methods
                1548-7091
                1548-7105
                6 July 2016
                25 July 2016
                September 2016
                01 March 2017
                : 13
                : 9
                : 751-754
                Affiliations
                [1 ]School of Life Sciences, University of Nottingham, Nottingham, UK
                Author notes
                [2 ]Correspondence to matt.loose@ 123456nottingham.ac.uk @mattloose
                Article
                EMS69060
                10.1038/nmeth.3930
                5008457
                27454285
                363b19a5-0f79-41c6-99f7-4d37f4a4381a

                Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use: http://www.nature.com/authors/editorial_policies/license.html#terms

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                Life sciences
                Life sciences

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