HIGH THROUGHPUT NUCLEIC ACID SEQUENCING BY EXPANSION AND RELATED METHODS

摘要

Nucleic acid sequencing methods and related products and methods for detection and presentation of the same are disclosed. Methods for sequencing a target nucleic acid comprise providing a daughter strand produced by a template-directed synthesis, the daughter strand comprising a plurality of subunits coupled in a sequence corresponding to a contiguous nucleotide sequence of all or a portion of the target nucleic acid, wherein the individual subunits comprise a tether, at least one probe or nucleobase residue, and at least one selectively cleavable bond. The selectively cleavable bond(s) is/are cleaved to yield a surrogate polymer of a length longer than the plurality of the subunits of the daughter strand, the surrogate polymer comprising the tethers and reporter elements for parsing genetic information in a sequence corresponding to the contiguous nucleotide sequence of all or a portion of the target nucleic acid. Reporter elements of the surrogate polymer are then detected. Disclosed methods for detecting the surrogate polymers comprise nanopore detection and other detection methods suitable for high-throughput DNA sequencing. Methods for presenting the surrogate polymer to the detector comprise presenting the surrogate polymers: 1) in flow, 2) tethered to a solid support, and 3) aligned on a substrate surface. Corresponding products, including surrogate polymers and oligomeric and monomeric substrate constructs are also disclosed.