GENERATING CAPTURE PROBES FOR SPATIAL ANALYSIS
The present disclosure relates to compositions and methods for generating capture probes on a substrate for identifying the location of analytes in a biological sample.
1 . A method of generating a spatial array comprising:
(a) immobilizing a first plurality of primers and a second plurality of primers on a substrate, wherein:
(i) a first primer of the first plurality of primers comprises one or more functional domains and a sequence complementary to a hybridization domain, wherein the one or more functional domains of the first primer comprise a sequencing specific site or a primer binding site; and
(ii) a second primer of the second plurality of primers comprises a sequence complementary to a capture domain;
(b) providing a template oligonucleotide comprising the hybridization domain capable of hybridizing to the sequence complementary to the hybridization domain of the first primer, a complement of a spatial barcode, and a complement of the capture domain;
(c) hybridizing the hybridization domain of the template oligonucleotide to the sequence complementary to the hybridization domain of the first primer;
(d) extending the first primer using the template oligonucleotide as a template to generate a capture probe comprising the first primer, the spatial barcode, and the capture domain; and
(e) releasing the template oligonucleotide from the capture probe, thereby generating the spatial array.