| CPC C12Q 1/6876 (2013.01) [C12Q 1/25 (2013.01); C12Q 1/6811 (2013.01); C12Q 1/6816 (2013.01); C12Q 1/6841 (2013.01); C12Q 1/6851 (2013.01)] | 30 Claims |
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1. A method for determining a location of a target nucleic acid in a biological sample comprising:
(a) providing the biological sample placed on a first substrate;
(b) contacting a plurality of first probes and second probes to the biological sample, wherein the plurality of first probes and second probes target a plurality of nucleic acids in the biological sample, wherein a first probe and a second probe of the plurality comprise sequences that are substantially complementary to sequences that are not adjacent to each other on the target nucleic acid, and wherein the second probe comprises a capture probe capture domain sequence that is complementary to all or a portion of a capture domain of a capture probe;
(c) hybridizing the first probe and the second probe to the target nucleic acid;
(d) generating a ligation product by ligating the first probe and the second probe;
(e) releasing the ligation product from the target nucleic acid;
(f) hybridizing the ligation product to the capture domain of the capture probe affixed on an array, wherein the capture probe further comprises a spatial barcode; and
(g) determining (i) all or part of the sequence of the ligation product bound to the capture domain, 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 determine the location of the target nucleic acid in the biological sample.
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