Methods for next generation genome walking and related compositions and kits
US10988802B2 · kind B2 · utility
Assignee
Inventor
Key dates
| Filing date | May 20, 2016 |
| Grant date | Apr 27, 2021 |
| Priority date | — |
| Expiry date | Feb 24, 2037 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q2600/16
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
Methods are provided herein for identifying rare and/or unknown DNA sequences by next-generation sequencing approaches. Isolated double-stranded (ds), single-stranded (ss), or ds/ss DNA is fragmented and the fragments are polished, phosphorylated, and tailed, as necessary. Fragmentation can be enzymatic or mechanical. A universal adapter sequence is ligated to each fragment, wherein the adapter can have a top strand without a 5′ phosphate, a 3′ with an —H in place of the —OH, and/or a 3′ extra base complementary to any base added to the polished fragments. The ligatamers may then serve as templates for amplification using a forward primer complementary to the adapter sequence and a reverse primer targeted to the fragment sequence. Compositions produced by these methods and kits adapted for performing these methods are also described herein.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.