Recombinase polymerase amplification

Details Recombinase polymerase amplification Recombinase polymerase amplification

C - Chemistry, Metallurgy

12

P

C12P 19/34 (2006.01) C12N 15/09 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2476481

This disclosure describes three related novel methods for Recombinase- Polymerase Amplification (RPA) of a target DNA that exploit the properties of the bacterial RecA and related proteins, to invade double-stranded DNA with single stranded homologous DNA permitting sequence specific priming of DNA polymerase reactions. The disclosed method has the advantage of not requiring tehrmocycling or thermophilic enzymes. Further, the improved processivity of the dislosed methods allow amplification of DNA up to hundreds of megabases in length.

L'invention concerne trois nouvelles méthodes d'amplification de la recombinase-polymérase (RPA) d'un ADN cible, utilisant les propriétés des protéines bactérienne RecA et analogues, pour envahir un ADN bicaténaire avec un ADN homologue monocaténaire permettant l'amorçage séquence-spécifique de réactions de l'ADN polymérase. La méthode de l'invention ne nécessite pas de thermocyclage ni d'enzymes thermophiles. Par ailleurs, ces nouvelles méthodes permettent d'amplifier l'ADN de plusieurs centaines de mégabases en longueur.

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