Patent Document ID: 10106789
Application ID: 15431660
Patent Flag: 1

Claim One:
1. A method for creating a library of polynucleotides, comprising the steps of: introducing multiple sets of initial probes into a plurality of compartments under conditions selected such that more than a half of the plurality of compartments contain one or less than one set of the initial probes, wherein each set of the initial probes comprises (1) an initial forward probe, wherein the initial forward probe (i) is affixed to a bead or a solid surface, and (ii) comprises a sequence complementary to a first subsequence of a first target sequence and one of at least 1,000 unique barcode sequences, and (2) an initial reverse probe, wherein the initial reverse probe comprises (i) a sequence complementary to a second subsequence of the first target sequence and (ii) a sequence complementary to a non-human, exogenous sequence; introducing multiple sets of second probes into the plurality of compartments, wherein each set of the second probes comprises (1) a second forward probe, wherein the second forward probe comprises (i) the non-human, exogenous sequence and (ii) a sequence that is complementary to a first subsequence of a second target sequence, and (2) a second reverse probe comprising a sequence that is complementary to a second subsequence to the second target sequence; amplifying the first target sequence using the multiple sets of the initial probes; amplifying the second target sequence using the multiple sets of the second probes; hybridizing the non-human exogenous sequence to its complement; and amplifying a fused sequence comprising one of the at least 1,000 barcode sequences, the first target sequence and the second target sequence, thereby generating a library of fused polynucleotides, wherein each of the fused polynucleotides comprises one of the at least 1,000 unique barcode sequences.