Patent ID: 11891654
Assignee: 10X GENOMICS, INC.
Field: Biotechnology (Chemistry)
Classification: CPC C | IPC C

Claim 0:
1. A method of identifying a location of a target nucleic acid in a permeabilized tissue section, the method comprising:
(a) generating a cDNA molecule comprising a sequence that is substantially complementary to the target nucleic acid using a reverse transcription primer comprising (i) a sequence that is substantially complementary to a portion of the target nucleic acid and (ii) a first adaptor sequence, wherein the step of generating the cDNA molecule occurs within the permeabilized tissue section;
(b) ligating a second adaptor sequence to a 3′ end of the cDNA molecule, wherein the step of ligating is performed within the tissue section;
(c) releasing the cDNA molecule ligated to the second adaptor sequence from the target nucleic acid in the tissue section, such that the second adaptor sequence contacts an array, wherein the array comprises an attached capture probe comprising in a 5′ to a 3′ direction: (i) a spatial barcode and (ii) a capture domain that hybridizes to the second adaptor sequence ligated to the cDNA molecule;
(d) extending a 3′ end of the capture probe using the cDNA molecule as a template; and
(e) determining (i) all or a part of the sequence of the target nucleic acid, or a complement thereof, and (ii) the sequence of the spatial barcode, or a complement thereof, and using the determined sequences of (i) and (ii) to identify the location of the target nucleic acid in the permeabilized tissue section.