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US 2017/0362647 A1Published application
Visualizing modified nucleotides and nucleic acid interactions in single cells
AssigneeSigma-Aldrich Co.
InventorsVikas Palhan, Carol Kreader
Life of this patent
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Abstract
Methods for visualizing modified nucleotides in a specific nucleic acid sequence or specific nucleic acid sequence interactions in single cells, wherein the methods comprise coupling an in situ hybridization (ISH) reaction with a proximity ligation as say (PLA) reaction.
Claims14
A method for visualizing a modified nucleotide in a specific nucleic acid sequence in a cell, the method comprising:
- a) contacting a prepared cell with at least one nucleic acid probe labeled with at least one label and having complementarity to the specific nucleic acid sequence, wherein the nucleic acid probe hybridizes with the specific nucleic acid sequence to form a hybridized cell;
- b) contacting the hybridized cell with a first binding agent that binds to the label of the nucleic acid probe and a second binding agent that binds to the modified nucleotide;
- c) detecting the first and second binding agents by a proximity ligation assay to visualize the modified nucleotide in the specific nucleic acid sequence in the cell.
A method for visualizing an interaction between two nucleic acid sequences in a cell, the method comprising:
- a) contacting a prepared cell with a first nucleic acid probe comprising a first label and having complementarity to a first nucleic acid sequence and a second nucleic acid probe comprising a second label and having complementarity to a second nucleic acid sequence, wherein the first and second labels are different and the first and second nucleic acid probes hybridize with the first and second nucleic acid sequences, respectively;
- b) contacting the cell from step a) with a first binding agent that binds to the first label and a second binding agent that binds to the second label;
- c) detecting the first and second binding agents by a proximity ligation assay to visualize the interaction between the two nucleic acid sequences in the cell.
Dependencies are inferred from the claim text.