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US 2022/0017892 A1Published application
Tiered ligation oligos
AssigneeFluent Biosciences Inc.
InventorsRobert Meltzer, Kristina Fontanez, Yi Xue
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Abstract
The disclosure provides methods for creating long oligonucleotide reagents that include barcodes and other element for sequencing library preparation, where the oligonucleotides are created by multiple tiers of ligation of shorter oligos. The disclosed methods work to extend short oligos that are attached to particles, thereby allowing one to create particles that carry large number of long sample preparation oligonucleotides without being required to synthesize those full-length molecules with a polymerase.
Claims19
A method for creating a target capture reagent, the method comprising:
- dividing a plurality of initial oligos into a set of partitions;
- ligating partition-specific first barcodes to the initial oligos to form ligation products;
- pooling the ligation products into a pool;
- splitting the pool into a second set of partitions;
- and
- ligating partition-specific second barcodes to the ligation products to form tripartite oligonucleotides each comprising (i) one of the initial oligos, (ii) one of the first barcodes, and (iii) one of the second barcodes.
3 more elements
Dependencies are inferred from the claim text.