Method and apparatus for simultaneous targeted sequencing of DNA, RNA and protein
US11365441B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | May 22, 2020 |
| Grant date | Jun 21, 2022 |
| Priority date | — |
| Expiry date | Aug 4, 2040 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q2521/537
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
Provided herein are methods and systems for the simultaneous targeted detection and sequencing of DNA, RNA, and Protein. In typical embodiments, the DNA, RNA, and Proteins are detected, characterized, and sequenced using just a single mammalian cell. One embodiment of detecting and characterizing DNA, RNA, or protein from a mammalian cell includes encapsulating a single cell in a drop and performing a protease digest on the encapsulated cell drop, performing a reverse transcriptase reaction; performing a droplet merger with barcoding PCR reagents and barcoding beads; performing a PCR reaction to attach the cell barcodes to the DNA targeted amplicons, RNA targeted amplicons, and protein tag amplicons, where all amplicons from the same emulsion contain the same cell barcode; and detecting and characterizing a DNA, RNA, or protein amplicon by sequencing the cell barcode incorporated into each amplicon.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.