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US 11,236,387 B2Granted
Compositions and methods for nucleic acid sequencing
AssigneeSingular Genomics Systems, Inc.
InventorsEli N. Glezer, Mohammad Vatankhah Varnosfaderani, Daan Witters, Vahid Karimkhani
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Abstract
Provided herein are methods and compositions for improved sequencing techniques using, for example, polymeric particles and/or three-dimensional structures.
Claims30
A method of amplifying a target polynucleotide, the method comprising:
- (a) contacting a composition comprising a plurality of cores with a sample comprising a target polynucleotide, wherein (i) each core of the plurality of cores is surrounded by a shell polymer, (ii) each core is formed by polymerized units of core monomers forming a core polymer, (iii) a core polynucleotide primer is attached to the core polymer within each core, (iv) the shell polymer is formed by polymerized units of shell monomers, and (v) the shell polymer is not attached to a polynucleotide primer;
- (b) amplifying the target polynucleotide to produce an amplicon, wherein amplifying comprises extension of the core primer hybridized to the target polynucleotide within each core.
A method of sequencing target polynucleotides, the method comprising:
- (a) contacting a plurality of particles with a sample comprising target polynucleotides, wherein each particle comprises a polymer covalently attached to polynucleotide primers;
- (b) amplifying the target polynucleotides to produce discrete amplicon clusters, wherein (i) amplifying comprises extension of the primers along the target polynucleotides, (ii) each amplicon cluster originates from amplification of a single target polynucleotide, and (iii) the amplicon clusters are arranged in multiple two-dimensional planes;
- and
- (c) sequencing the amplicon clusters, wherein sequencing comprises detecting sequences of signals from the particles through each of the plurality of two-dimensional planes;
- wherein each particle comprises a core surrounded by a shell polymer, wherein:
- (a) each core is formed by polymerized units of core monomers forming a core polymer;
- (b) a core polynucleotide primer is attached to the core polymer within each core;
- (c) a target nucleic acid is capable of hybridizing to the core polynucleotide primer;
- (d) the shell polymer is formed by polymerized units of shell monomers;
- and
- (e) the shell polymer is not attached to a polynucleotide primer.
8 more elements
Dependencies are inferred from the claim text.