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

Claim 1:
2. 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) extending a 3′ end of the cDNA molecule to include a second adaptor sequence thereby generating an extended cDNA molecule, wherein the step of extending is performed within the tissue section;
(c) releasing the extended cDNA molecule from the target nucleic acid, such that the extended cDNA molecule 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 binds to the second adaptor sequence of the extended cDNA molecule;
(d) extending a 3′ end of the capture probe using the extended 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.