Kinetic exclusion amplification of nucleic acid libraries
First Claim
1. A method for determining nucleic acid sequences, comprising performing a sequencing procedure that detects an apparently clonal population of amplicons at each of a plurality of amplicon sites on an array, wherein the array is made by a process that comprises(a) providing an amplification reagent comprising(i) a plurality 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 the amplification sites,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 several nucleic acids in the plurality of different nucleic acids;
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(i) transporting the different target nucleic acids from the solution to the amplification sites, and(ii) amplifying the target nucleic acids that are at the amplification sites at an amplification rate that promotes filling the capacity of the amplification sites and exclusion of more than one of the different target nucleic acids from attaching at each amplification site to produce an array of amplicon sites that each comprise the apparently clonal population of amplicons.
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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.
49 Citations
25 Claims
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1. A method for determining nucleic acid sequences, comprising performing a sequencing procedure that detects an apparently clonal population of amplicons at each of a plurality of amplicon sites on an array, wherein the array is made by a process that comprises
(a) providing an amplification reagent comprising (i) a plurality 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 the amplification sites, 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 several nucleic acids in the plurality of different nucleic acids; - and
(b) reacting the amplification reagent, wherein the plurality of different target nucleic acids in the solution is at a concentration that results in simultaneously; (i) transporting the different target nucleic acids from the solution to the amplification sites, and (ii) amplifying the target nucleic acids that are at the amplification sites at an amplification rate that promotes filling the capacity of the amplification sites and exclusion of more than one of the different target nucleic acids from attaching at each amplification site to produce an array of amplicon sites that each comprise the apparently clonal population of amplicons. - 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)
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Specification