Enrichment of DNA sequencing libraries from samples containing small amounts of target DNA
US10576446B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | May 2, 2014 |
| Grant date | Mar 3, 2020 |
| Priority date | — |
| Expiry date | May 2, 2034 |
Classification
- Technology area (CPC B)Performing Operations; Transporting
- CPC primaryB01J2219/00722
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
Provided herein is a method for capturing DNA molecules in solution. The method may comprise: extracting DNA from a sample that comprises endogenous DNA and environmental DNA to produce extracted DNA; ligating universal adaptors to the extracted DNA; hybridizing the extracted DNA, in solution, with affinity-tagged RNA probes generated by: in vitro transcribing a library of fragmented reference genomic DNA that has been ligated to an RNA promoter adaptor, in the presence of an affinity-tagged ribonucleotide; binding the product with a capture agent that is tethered to a substrate in the presence of RNA oligonucleotides that are complementary to the adaptors, thereby capturing the hybridized DNA molecules on the substrate; washing the substrate to remove any unbound DNA molecules; and releasing the captured DNA molecules. A kit for performing the method is also provided.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.