Patent ID: 7842457

Claim:
A method for amplifying one or more nucleic acids onto a bead comprising the steps of: (a) forming a water-in-oil emulsion to create a plurality of aqueous microreactors wherein a plurality of the microreactors comprises on average one single stranded nucleic acid template, a single bead with a first population comprising a plurality of molecules of a first primer species disposed thereon, and an amplification reaction solution comprising a second population comprising a plurality of molecules of the first primer species and a plurality of molecules of a second primer species and reagents necessary to perform nucleic acid amplification, wherein the first primer species is capable of binding to the single stranded nucleic acid template, the second primer species is capable of binding to a complementary strand of the single stranded nucleic acid template, and further wherein the molecules of the second primer species and the molecules of the first population of the primer species are each present in greater numbers within the aqueous microreactors than the number of molecules of the second population of the first primer species; (b) asymmetrically amplifying the single stranded nucleic acid template and the complementary strand to the template strand in the amplification reaction solution to form a population of amplified copies of the single stranded template nucleic acid, wherein substantially all of the molecules of the second population of the first primer species in the amplification reaction solution are depleted; (c) binding a plurality of the asymmetrically amplified copies of the single stranded template nucleic acid to the first population of the first primer species on the bead in a plurality of the microreactors, wherein at least 100,000 bead bound complementary strands are extended from the first primer species to form a population of beads with amplified nucleic acid template bound thereto; (d) breaking the emulsion to release the population of beads with amplified nucleic acid template bound thereto from the microreactors and away from the amplification reaction solution comprising unbound amplification products; and (e) enriching for beads with amplified nucleic acid template bound thereto by removing beads to which no nucleic acid is bound; and (f) distributing the beads with amplified nucleic acid template bound thereto onto an array.