US 11,667,957 B2
Methods and compositions for identifying ligands on arrays using indexes and barcodes
Darren Segale, San Diego, CA (US); Fiona E. Black, Encinitas, CA (US); Jeffrey Dennis Brodin, San Diego, CA (US); Jeffrey Fisher, San Diego, CA (US); Lorenzo Berti, San Diego, CA (US); and Siew Hong Leong, Singapore (SG)
Assigned to Illumina, Inc., San Diego, CA (US)
Filed by Illumina, Inc., San Diego, CA (US)
Filed on Oct. 23, 2019, as Appl. No. 16/661,885.
Claims priority of provisional application 62/903,108, filed on Sep. 20, 2019.
Claims priority of provisional application 62/750,370, filed on Oct. 25, 2018.
Prior Publication US 2020/0131570 A1, Apr. 30, 2020
Int. Cl. C12Q 1/6816 (2018.01); C12Q 1/6837 (2018.01)
CPC C12Q 1/6816 (2013.01) 20 Claims
OG exemplary drawing
 
1. A method for identifying target ligands in an array comprising:
(a) obtaining a first and a second subpopulation of beads, wherein each bead comprises:
(i) a capture probe capable of specifically binding to a target ligand,
(ii) a first polynucleotide comprising a barcode, wherein the barcode is indicative of the capture probe, and
(iii) a second polynucleotide comprising an index, wherein each index of the first subpopulation of beads is different from each index of the second subpopulation of beads;
(b) specifically binding first target ligands to the capture probes of the first subpopulation of beads in a first reaction volume, and specifically binding second target ligands to the capture probes of the second subpopulation of beads in a second reaction volume; and
(c) distributing the first and the second subpopulations of beads comprising the specifically bound first and second target ligands on a surface of a substrate, thereby obtaining an array of beads;
(d) detecting in the array the specifically bound first and second target ligands of the first and the second subpopulations of beads;
(e) sequencing the barcodes in the array of beads;
(f) sequencing the indexes in the array of beads; and
(g) decoding the locations of beads in the array of beads comprising a detected target ligand in the array, thereby identifying target ligands in the array of beads.