Abstract
Nanopore DNA sequencing is a single molecule sequencing technique with promise for long reads, low cost, and minimal sample preparation. We have built on our progress in nucleotide sensitivity and DNA control to interpret the procession of ion current levels observed as DNA translocates through the nanopore MspA. Approximately 4 nucleotides affect the ion current of each level, therefore, we measured current corresponding to all 256 4-nucleotide combinations (quadromers). This quadromer map proves to be highly predictive of ion current levels for nanopore reads of bacteriophage phi X 174. We show nanopore sequencing reads of up to 4,500 bases in length, which can be unambiguously aligned the phi X 174 reference genome. We also demonstrate proof of concept hybrid genome assembly and polymorphism detection.
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