Kinetic exclusion amplification of nucleic acid libraries
First Claim
1. A method for amplifying nucleic acids, comprising(a) providing an amplification reagent comprising(i) an array of amplification sites, and(ii) a solution comprising a plurality of different target nucleic acids,wherein the number of the different target nucleic acids in the solution exceeds the number of amplification sites in the array,wherein the different target nucleic acids have fluidic access to the plurality of amplification sites, andwherein each of the amplification sites comprises a capacity for more than one of the nucleic acids in the plurality of different nucleic acids;
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Abstract
A method including (a) providing an amplification reagent including an array of sites, and a solution having different target nucleic acids; and (b) reacting the amplification reagent to produce amplification sites each having a clonal population of amplicons from a target nucleic acid from the solution. The reacting can include simultaneously transporting the nucleic acids to the sites at an average transport rate, and amplifying the nucleic acids that transport to the sites at an average amplification rate, wherein the average amplification rate exceeds the average transport rate. The reacting can include producing a first amplicon from a nucleic acid that transports to each of the sites, and producing subsequent amplicons from the nucleic acid or from the first amplicon, wherein the average rate at which the subsequent amplicons are generated exceeds the average rate at which the first amplicon is generated.
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Citations
28 Claims
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1. A method for amplifying nucleic acids, comprising
(a) providing an amplification reagent comprising (i) an array of amplification sites, and (ii) a solution comprising a plurality of different target nucleic acids, wherein the number of the different target nucleic acids in the solution exceeds the number of amplification sites in the array, wherein the different target nucleic acids have fluidic access to the plurality of amplification sites, and wherein each of the amplification sites comprises a capacity for more than one of the nucleic acids in the plurality of different nucleic acids; - and
(b) reacting the amplification reagent to produce a plurality of amplification sites that each comprise a clonal population of amplicons from an individual target nucleic acid from the solution, wherein the plurality of different target nucleic acids is at a concentration that results in simultaneously (i) transporting the different target nucleic acids to attach at the amplification sites, and (ii) amplifying the target nucleic acids that are at the amplification sites, wherein the amplifying at each amplification site occurs at a rate to promote filling the capacity of the amplification site and exclusion of more than one of the different target nucleic acids from attaching at said each amplification site. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28)
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Specification