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US 10,138,509 B2Granted
Method for generating a three-dimensional nucleic acid containing matrix
AssigneePresident and Fellows of Harvard College
InventorsGeorge M. Church, Je-Hyuk Lee, Richard C. Terry, Evan R. Daugharthy
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Abstract
Methods of making a three-dimensional matrix of nucleic acids within a cell is provided.
Claims43
A method of identifying nucleic acids within a cell, comprising:
- contacting a plurality of nucleic acids having a relative three-dimensional spatial relationship within the cell with a matrix-forming material in a manner to substantially retain the relative three-dimensional spatial relationship;
- using the matrix-forming material to form a three-dimensional polymerized matrix including the nucleic acids of the plurality of nucleic acids covalently bound to the three-dimensional polymerized matrix;
- and
- detecting signals from the nucleic acids or derivatives thereof, thereby identifying the nucleic acids.
1 more element
A method of identifying one or more nucleic acids within a cell comprising:
- contacting a plurality of nucleic acids having a relative three-dimensional spatial relationship within the cell with a matrix-forming material to form a three-dimensional polymerized matrix comprising the plurality of nucleic acids coupled to the three-dimensional polymerized matrix, wherein the relative three-dimensional spatial relationship is substantially retained in the three-dimensional polymerized matrix;
- amplifying the plurality of nucleic acids to produce amplicons within the three-dimensional polymerized matrix;
- coupling the amplicons to the three-dimensional polymerized matrix;
- and
- detecting the amplicons to identify a sequence of the one or more nucleic acids.
2 more elements
A method of identifying a relative three-dimensional spatial relationship of one or more nucleic acids within a cell comprising:
- contacting a plurality of nucleic acids having the relative three-dimensional spatial relationship within the cell with a matrix-forming material in a manner to substantially retain the relative three-dimensional spatial relationship;
- using the matrix-forming material to form a three-dimensional polymerized matrix including the plurality of nucleic acids covalently bound to the three-dimensional polymerized matrix;
- amplifying the plurality of nucleic acids to produce amplicons within the matrix, covalently bonding the amplicons to the three-dimensional polymerized matrix;
- labeling the amplicons with a detectable label;
- and
- imaging the amplicons to identify the relative three-dimensional spatial relationship of the one or more nucleic acids within the cell.
3 more elements
Dependencies are inferred from the claim text.