Patent ID: 7485428

Claim:
A method of recombinase-polymerase amplification of a double-stranded target sequence polymer molecule, comprising: (a) contacting a recombinase agent with a first primer sequence molecule to form a first recombinase agent-primer pair and contacting a recombinase agent with a second primer sequence molecule to form a second recombinase agent-primer pair; (b) contacting the first and second recombinase agent-primer pairs with the double-stranded target sequence polymer molecule to form a structure, wherein the first and second primer sequence molecules of the recombinase agent-primer pairs associate with the double-stranded target sequence polymer molecule; (c) extending the first and second primer sequence molecules along the double-stranded target sequence polymer molecule with a polymerase molecule and monomers to be incorporated into the extended first and second primer sequence molecules, using the double-stranded target sequence polymer molecule as a template; (d) contacting the double-stranded target sequence polymer molecule with primosome assembly proteins, a clamp loader/polymerase III holoenzyme complex, a DNA polymerase III core, a DNA polymerase I, a primase, a helicase, and a ligase; (e) replicating the strand of the double-stranded target sequence polymer molecule that is displaced by the recombinase agent-primer pair structures through lagging strand synthesis; and (f) repeating steps (b) through (e) until a desired degree of amplification is achieved.