Recombinase polymerase amplification
US7666598B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Apr 11, 2005 |
| Grant date | Feb 23, 2010 |
| Priority date | — |
| Expiry date | May 26, 2025 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q1/70
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
This disclosure describes related novel methods for Recombinase-Polymerase Amplification (RPA) of a target DNA that exploit the properties of recombinase and related proteins, to invade double-stranded DNA with single stranded homologous DNA permitting sequence specific priming of DNA polymerase reactions. The disclosed methods have the advantage of not requiring thermocycling or thermophilic enzymes, thus offering easy and affordable implementation and portability relative to other amplification methods. Further disclosed are conditions to enable real-time monitoring of RPA reactions, methods to regulate RPA reactions using light and otherwise, methods to determine the nature of amplified species without a need for gel electrophoresis, methods to improve and optimize signal to noise ratios in RPA reactions, methods to optimize oligonucleotide primer function, methods to control carryover contamination, and methods to employ sequence-specific third ‘specificity’ probes. Further described are novel properties and approaches for use of probes monitored by light in dynamic recombination environments.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.