Single nucleotide detection method and associated probes
US11920192B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | May 15, 2018 |
| Grant date | Mar 5, 2024 |
| Priority date | — |
| Expiry date | May 25, 2039 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q2563/159
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
A method of sequencing a nucleic acid comprising (1) generating a stream of single nucleoside triphosphates by progressive enzymatic digestion of the nucleic acid; (2) producing at least one oligonucleotide used probe by reacting, in the presence of a polymerase, at least one of the single nucleoside triphosphates with a corresponding biological probe comprising (a) a first single-stranded oligonucleotide including an exonuclease blocking-site, a restriction endonuclease recognition-site located on the 5′ side of the blocking-site and including a single nucleotide capture-site e, and at least one fluorophore region and (b) a second and optionally a third single-stranded oligonucleotide each separate from the first oligonucleotide; (3) cleaving the first oligonucleotide strand of the used probe at the recognition-site with a restriction endonuclease; (4) digesting the first oligonucleotide component with an enzyme to yield fluorophores in a detectable state and (5) detecting the fluorophores released in step (4).
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.