Multiplex targeted amplification using flap nuclease
US7862999B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Jan 17, 2008 |
| Grant date | Jan 4, 2011 |
| Priority date | — |
| Expiry date | Jun 29, 2028 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q1/6853
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
Methods for multiplex amplification of a plurality of targets of distinct sequence from a complex mixture are disclosed. In one aspect targets are circularized using a single circularization probe that is complementary to two regions in the target that flank a region to be amplified. The targets may hybridize to the circularization probe so that 5′ or 3′ flaps are generated and methods for removing flaps and circularizing the resulting product are disclosed. In another aspect targets are hybridized to dU probes so that 5′ and 3′ flaps are generated. The flaps are cleaved using 5′ or 3′ flap endonucleases or 3′ to 5′ exonucleases. The target sequences are then ligated to common primers, the dU probes digested and the ligated targets amplified.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.