Single nucleotide detection method
US9856528B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Jul 22, 2015 |
| Grant date | Jan 2, 2018 |
| Priority date | — |
| Expiry date | Jul 22, 2035 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q2565/629
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
A method of sequencing a nucleic acid involves (1) generating single nucleoside triphosphates by progressive pyrophosphorolysis of the nucleic acid; (2) producing a substantially double-stranded oligonucleotide used probe by reacting one of the single nucleoside triphosphates with a corresponding probe system comprising (a) a first single-stranded oligonucleotide labelled with characteristic detectable elements in an undetectable state and (b) second and third single-stranded oligonucleotides capable of hybridising to complementary regions on the first oligonucleotide; (3) digesting the used probe with an enzyme having double-stranded exonucleolytic activity to yield the detectable elements in a detectable state and a single-stranded fourth oligonucleotide which is at least in part the complement of the first oligonucleotide; (4) reacting the fourth oligonucleotide with another first oligonucleotide to produce a substantially double-stranded oligonucleotide product corresponding to the used probe; (5) repeating (3) and (4) in a cycle and (6) detecting the detectable elements released in each iteration.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.