US 12,286,667 B2
In situ detection of nucleic acid molecules
Eli N. Glezer, Del Mar, CA (US); Hu Cang, San Diego, CA (US); and Zhenmin Hong, San Diego, CA (US)
Assigned to SINGULAR GENOMICS SYSTEMS, INC., San Diego, CA (US)
Filed by Singular Genomics Systems, Inc., San Diego, CA (US)
Filed on Sep. 19, 2023, as Appl. No. 18/469,871.
Application 18/469,871 is a continuation of application No. 17/938,329, filed on Oct. 5, 2022.
Application 17/938,329 is a continuation of application No. 17/816,841, filed on Aug. 2, 2022.
Application 17/816,841 is a continuation of application No. 17/840,473, filed on Jun. 14, 2022.
Application 17/840,473 is a continuation of application No. 17/840,469, filed on Jun. 14, 2022, granted, now 11,643,679, issued on May 9, 2023.
Application 17/840,469 is a continuation of application No. 17/396,575, filed on Aug. 6, 2021, granted, now 11,492,662, issued on Nov. 8, 2022.
Claims priority of provisional application 63/062,054, filed on Aug. 6, 2020.
Claims priority of provisional application 63/140,700, filed on Jan. 22, 2021.
Claims priority of provisional application 63/209,886, filed on Jun. 11, 2021.
Prior Publication US 2024/0026428 A1, Jan. 25, 2024
This patent is subject to a terminal disclaimer.
Int. Cl. C12Q 1/6841 (2018.01); C12Q 1/6804 (2018.01); G01N 33/53 (2006.01)
CPC C12Q 1/6841 (2013.01) [C12Q 1/6804 (2013.01); G01N 33/5308 (2013.01); G01N 33/53 (2013.01)] 27 Claims
 
1. A method of detecting nucleic acid molecules in a sample comprising a cell or tissue, said method comprising:
i) contacting the sample with a first polynucleotide probe and binding said first polynucleotide probe to a first nucleic acid molecule, and contacting the sample with a second polynucleotide probe and binding said second polynucleotide probe to a second nucleic acid molecule, wherein said first polynucleotide probe comprises a first oligonucleotide binding sequence and a first sequence, and wherein said second polynucleotide probe comprises a second oligonucleotide binding sequence and a second sequence;
ii) amplifying said first and second polynucleotide probes to generate amplification products; and
iii) hybridizing a first oligonucleotide to the first oligonucleotide binding sequence, and detecting a series of fluorescent signals associated with the first sequence; followed by hybridizing the second oligonucleotide to the second oligonucleotide binding sequence, and detecting a series of fluorescent signals associated with the second sequence, wherein the first oligonucleotide and the second oligonucleotide comprise different sequences.