POLYNUCLEOTIDE BARCODE GENERATION
The present disclosure provides compositions, methods, systems, and devices for polynucleotide processing. Such polynucleotide processing may be useful for a variety of applications, including polynucleotide sequencing. In some cases, this disclosure provides methods for the generation of polynucleotide barcode libraries, and for the attachment of such polynucleotides to target polynucleotides.
1 .- 62 . (canceled)
63 . A library, comprising:
a plurality of nucleic acid barcode sequences comprising at least 1000 different nucleic acid barcode sequences;
a plurality of partitions comprising at least 1000 partitions, wherein each of at least 1000 different nucleic acid barcode sequences in the plurality of barcode sequences is disposed within a separate partition; and
wherein each of at least 1000 partitions of the plurality of partitions comprises more than 1000 nucleic acid barcode molecules comprising the same nucleic acid barcode sequence.
64 . The library of claim 63 , wherein the nucleic acid barcode sequences comprise 4 or more nucleotides in length.
65 . The library of claim 63 , wherein the nucleic acid barcode sequences are from about 4 to about 19 nucleotides in length.
66 . The library of claim 63 , wherein the plurality of nucleic acid barcode sequences comprises at least 5,000 different nucleic acid barcode sequences.
67 . The library of claim 66 , comprising at least 5,000 partitions, and each of at least 5,000 different nucleic acid barcode sequences in the plurality of barcode sequences is disposed within a separate partition.
68 . The library of claim 64 , wherein the plurality of nucleic acid barcode sequences comprises at least 10,000 different nucleic acid barcode sequences.
69 . The library of claim 64 , wherein the plurality of nucleic acid barcode sequences comprises at least 100,000 different nucleic acid barcode sequences.
70 . The library of claim 69 , comprising at least 100,000 partitions, and each of at least 100,000 different nucleic acid barcode sequences in the plurality of barcode sequences is disposed within a separate partition.
71 . The library of claim 63 , wherein each of the plurality of partitions comprises at least about 5,000 nucleic acid barcode molecules comprising the same nucleic acid barcode sequence.
72 . The library of claim 63 , wherein each of the plurality of partitions comprises at least about 10,000 nucleic acid barcode molecules comprising the same nucleic acid barcode sequence.
73 . The library of claim 63 , wherein each of the plurality of partitions comprises at least about 100,000 nucleic acid barcode molecules comprising the same nucleic acid barcode sequence.
74 . The library of claim 63 , wherein each of the plurality of partitions comprises at least about 200,000 nucleic acid barcode molecules comprising the same nucleic acid barcode sequence.
75 . The library of claim 63 , wherein each of the plurality of partitions comprises a different nucleic acid barcode sequence.
76 . The library of claim 63 , wherein two or more of the plurality of partitions comprise the same nucleic acid barcode sequence.
77 . The library of claim 63 , wherein the plurality of partitions are contained within one or more vessels, wherein at least one of the one or more vessels comprises at least 2 partitions.
78 . The library of claim 63 , wherein the plurality of partitions are contained within one or more vessels, where at least one of the one or more vessels comprises at least 1000 partitions.
79 . The library of claim 63 , wherein the plurality of partitions are contained within one or more vessels, where at least one of the one or more vessels comprises at least 10,000 partitions.
80 . The library of claim 63 , wherein the plurality of partitions are contained within one or more vessels, where at least one of the one or more vessels comprises at least 50,000 partitions.
81 . The library of claim 63 , wherein at least 1000 partitions of the plurality of partitions are contained in less than 1000 separate vessels.
82 . The library of claim 63 , wherein at least 1000 partitions of the plurality of partitions are contained in fewer than 10 separate vessels.
83 . The nucleic acid barcode library of claim 63 , wherein at least 1000 partitions of the plurality of partitions are contained in 2 or fewer separate vessels.
84 . The library of claim 63 , wherein the plurality of partitions are contained in 1 vessel.
85 . The library of claim 63 , wherein the plurality of partitions comprise capsules.
86 . The library of claim 63 , wherein the plurality of partitions comprise droplets in an emulsion.
87 . The library of claim 63 , wherein the plurality of partitions comprise capsules in droplets in an emulsion.
88 . The library of claim 63 , wherein the one or more vessels are selected from containers, tubes, wells and microwells.
89 . The library of claim 63 , wherein the partitions lack one or more components necessary to perform an amplification reaction.
90 . The library of claim 63 , wherein each barcode molecule further comprises at least one functional nucleic acid sequence.
91 . The library of claim 90 , wherein the at least one functional nucleic acid sequence is selected from a primer sequence, a primer annealing sequence, a polynucleotide immobilization sequence, and a ligation adapter sequence.
92 . The library of claim 91 , wherein the primer sequence comprises a MALBAC primer.