US 12,435,364 B2
Probes comprising a split barcode region and methods of use
Malte Kühnemund, Stockholm (SE)
Assigned to 10x Genomics, Inc., Pleasanton, CA (US)
Filed by 10x Genomics, Inc., Pleasanton, CA (US)
Filed on Aug. 15, 2022, as Appl. No. 17/888,361.
Claims priority of provisional application 63/233,599, filed on Aug. 16, 2021.
Prior Publication US 2023/0061542 A1, Mar. 2, 2023
Int. Cl. C12Q 1/6841 (2018.01); C12Q 1/6827 (2018.01); C12Q 1/6837 (2018.01); C12Q 1/6851 (2018.01); C12Q 1/6876 (2018.01)
CPC C12Q 1/6841 (2013.01) [C12Q 1/6827 (2013.01); C12Q 1/6837 (2013.01); C12Q 1/6876 (2013.01); C12Q 1/6851 (2013.01)] 20 Claims
OG exemplary drawing
 
1. A method for analyzing a biological sample, comprising:
(a) contacting the biological sample with:
(i) a first probe comprising a first hybridization region and a first portion of a barcode region, and
(ii) a second probe comprising a second hybridization region and a second portion of the barcode region,
wherein the first and second hybridization regions are complementary to target sequences in a target RNA molecule in the biological sample,
wherein the first probe and the second probe hybridize to the target RNA molecule in the biological sample, and
wherein the barcode region comprises one or more barcode sequences corresponding to the target RNA molecule;
(b) ligating the first and second probes hybridized to the target RNA molecule to form a composite probe by connecting the first and second portions of the barcode region and by connecting the ends of the first hybridization region and the second hybridization region; and
(c) contacting the biological sample with a detectable probe that hybridizes to the barcode region at sequences corresponding to both the first portion of the barcode region and the second portion of the barcode region, or a complement thereof,
wherein a signal associated with the detectable probe is detected in the biological sample, thereby detecting the target RNA molecule in the biological sample.