Patent · US Active

Recombinase polymerase amplification

US8030000B2 · kind B2 · utility

90Cited by
20References
23Claims
0Family size

Assignee

Inventors

Key dates

Filing dateFeb 19, 2010
Grant dateOct 4, 2011
Priority date
Expiry dateFeb 19, 2030

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 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.