US 12,006,534 B2
Spatial sequencing
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 Jul. 21, 2023, as Appl. No. 18/356,724.
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.
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. 2, 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 2023/0366013 A1, Nov. 16, 2023
Int. Cl. C12Q 1/68 (2018.01); C12Q 1/6841 (2018.01)
CPC C12Q 1/6841 (2013.01) 20 Claims
OG exemplary drawing
 
1. A method of detecting an RNA molecule in a cell or tissue, said method comprising:
contacting a cell or tissue with a polynucleotide probe and hybridizing a first hybridization sequence of the polynucleotide probe to a first target sequence of the RNA molecule, and hybridizing a second hybridization sequence of the polynucleotide probe to a second target sequence of the RNA molecule, wherein said RNA molecule comprises a target sequence between the first target sequence and the second target sequence;
extending the polynucleotide probe along the target sequence to generate a complement of the target sequence, and ligating the complement of the target sequence to the polynucleotide probe thereby forming a circular oligonucleotide;
extending an amplification primer hybridized to the circular oligonucleotide with a polymerase to generate an extension product comprising the target sequence; and
hybridizing a sequencing primer to the extension product and sequencing the target sequence in the cell or tissue, thereby detecting the RNA molecule.