Recombinase polymerase amplification
US9309502B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Jan 30, 2015 |
| Grant date | Apr 12, 2016 |
| Priority date | — |
| Expiry date | Jan 30, 2035 |
Classification
- Technology area (CPC B)Performing Operations; Transporting
- CPC primaryB01L2300/18
- WIPO fieldMeasurement
- WIPO sectorInstruments
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 carry-over 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.