4.5 Article

Error analysis of idealized nanopore sequencing

Journal

ELECTROPHORESIS
Volume 34, Issue 15, Pages 2137-2144

Publisher

WILEY-BLACKWELL
DOI: 10.1002/elps.201300174

Keywords

DNA sequencing; Error rates; Nanopore; Nucleic acids; Single-molecule sequencing

Funding

  1. National Institutes of Health [1R21HG006561-01]

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This numerical study provides an error analysis of an idealized nanopore sequencing method in which ionic current measurements are used to sequence intact single-stranded DNA in the pore, while an enzyme controls DNA motion. Examples of systematic channel errors when more than one nucleotide affects the current amplitude are detailed, which if present will persist regardless of coverage. Absent such errors, random errors associated with tracking through homopolymer regions are shown to necessitate reading known sequences (Escherichia coli K-12) at least 140 times to achieve 99.99% accuracy (Q40). By exploiting the ability to reread each strand at each pore in an array, arbitrary positioning on an error rate versus throughput tradeoff curve is possible if systematic errors are absent, with throughput governed by the number of pores in the array and the enzyme turnover rate.

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