Digital nuclease detection compositions and methods
US12378591B2 · kind B2 · utility
Assignee
Inventor
Key dates
| Filing date | Sep 27, 2018 |
| Grant date | Aug 5, 2025 |
| Priority date | — |
| Expiry date | Mar 7, 2042 |
Classification
- Technology area (CPC G)Physics
- CPC primaryG01N2333/922
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
In certain embodiments, the present invention provides a detection composition comprising a picodroplet comprising (a) an aqueous solution, and (b) a substrate probe comprising (i) an oligonucleotide of 2 to 75 nucleotides in length, (ii) a fluorophore operably linked to the oligonucleotide, and (iii) a quencher operably linked to the oligonucleotide. As used herein, the term “picodroplet” comprises a liquid droplet that has a volume of 0.014 to 2.6 picoliters. In certain embodiments, the present invention provides a method of detecting at least one individual nuclease molecule present in a sample, comprising contacting an aqueous sample suspected of containing at least one nuclease with at least one detection composition comprising a picodroplet comprising (a) an aqueous solution, and (b) a substrate probe comprising (i) an oligonucleotide of 2 to 75 nucleotides in length, (ii) a fluorophore operably linked to the oligonucleotide, and (iii) a quencher operably linked to the oligonucleotide to form an aqueous reaction mixture; emulsifying the aqueous mixture in oil to form picoliter-scale droplets in an emulsion, (c) incubating the picoliter-scale droplets in the emulsion in order …
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.