US 12,486,532 B2
Detecting molecules in tissues
Eli N. Glezer, Del Mar, CA (US)
Assigned to Singular Genomics Systems, Inc., San Diego, CA (US)
Filed by Singular Genomics Systems, Inc., San Diego, CA (US)
Filed on Apr. 24, 2024, as Appl. No. 18/645,232.
Application 18/645,232 is a continuation of application No. 18/356,724, filed on Jul. 21, 2023, granted, now 12,006,534.
Application 18/356,724 is a continuation of application No. 18/319,244, filed on May 17, 2023.
Application 18/319,244 is a continuation of application No. 18/045,913, filed on Oct. 12, 2022, granted, now 11,753,678, issued on Sep. 12, 2023.
Application 18/045,913 is a continuation of application No. 17/815,542, filed on Jul. 27, 2022, granted, now 11,680,288, issued on Jun. 20, 2023.
Application 17/815,542 is a continuation of application No. 17/396,579, filed on Aug. 6, 2021, granted, now 11,434,525, issued on Sep. 6, 2022.
Claims priority of provisional application 63/209,903, filed on Jun. 11, 2021.
Claims priority of provisional application 63/140,703, filed on Jan. 22, 2021.
Claims priority of provisional application 63/062,047, filed on Aug. 6, 2020.
Prior Publication US 2024/0287590 A1, Aug. 29, 2024
Int. Cl. C12Q 1/68 (2018.01); C12Q 1/6841 (2018.01)
CPC C12Q 1/6841 (2013.01) 25 Claims
 
1. A method of detecting molecules in a tissue, said method comprising:
(i) binding a first polynucleotide probe to a first nucleic acid molecule in said tissue and incorporating a sequence of the first nucleic acid molecule into the first polynucleotide probe; forming a first amplification product comprising multiple complements of the first polynucleotide probe; and binding a first primer to the first amplification product and incorporating a first fluorescently labeled nucleotide into the first primer;
(ii) binding a second polynucleotide probe to an oligonucleotide of an antibody-oligonucleotide conjugate, wherein said antibody-oligonucleotide conjugate is attached to a protein in said tissue; forming a second amplification product comprising multiple complements of the second polynucleotide probe; and binding a second primer to the second amplification product and incorporating a second fluorescently labeled nucleotide into the second primer;
(iii) contacting the tissue with a stain; and
(iv) detecting the first fluorescently labeled nucleotide, the second fluorescently labeled nucleotide, and the stain.