Methods for determining base locations in a polynucleotide
US10760117B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Apr 5, 2016 |
| Grant date | Sep 1, 2020 |
| Priority date | — |
| Expiry date | Apr 25, 2036 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q2565/631
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
Disclosed are methods for polynucleotide sequencing that detect the location of selected nucleobases with greater precision. The methods can be used to determine the location and nature of modified bases in a polynucleotide, that is, non-canonical bases, or to improve accuracy of sequencing of “problem” regions of DNA sequencing such as homopolymers, GC rich areas, etc. The sequencing method exemplified is nanopore sequencing. Nanopore sequencing is used to generate a unique signal at a point in a polynucleotide sequence where an abasic site (AP site, or apurinic or apyrimidinic site) exists. As part of the method, an abasic site is specifically created enzymatically using a DNA glycosylase that recognizes a pre-determined nucleobase species and cleaves the N-glycosidic bond to release only that base, leaving an AP site in its place.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.