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US 12,006,534 B2Granted
Spatial sequencing
AssigneeSingular Genomics Systems, Inc.
InventorEli N. Glezer
Life of this patent
Estimating the term…
Abstract
Disclosed herein, inter alia, are compositions and methods of use thereof for interrogating a sample comprising a cell.
Claims20
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.
2 more elements
A method of incorporating a modified nucleotide into a nucleic acid 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 an 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;
- amplifying the circular oligonucleotide by extending a first amplification primer hybridized to the circular oligonucleotide with a polymerase to generate an extension product comprising the target sequence;
- and
- hybridizing a primer to the extension product and incorporating a modified nucleotide into the primer, wherein the modified nucleotide comprises a blocking group capable of preventing polymerase extension.
2 more elements
Dependencies are inferred from the claim text.