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US 2013/0281306 A1Published application
Sequencing methods
AssigneeIllumina, Inc.
InventorsRoberto Rigatti, Jonathan Mark Boutell, Jason Richard Betley, Niall Anthony Gormley, Mostafa Ronaghi, Dirk Evers
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Abstract
The present technology relates to molecular sciences, such as genomics. More particularly, the present technology relates to nucleic acid sequencing.
Claims32
A method of generating overlapping sequencing reads, said method comprising:
- providing a substrate having a surface, said surface comprising a site having a plurality of polynucleotides attached thereto;
- and
- performing sequencing runs on said plurality of polynucleotides, said sequencing runs being initiated at different times such that said sequencing runs proceed for different durations, thereby generating a set of overlapping sequencing reads, wherein performing a sequencing run comprises:
- (a) providing a first sequencing reagent to a target nucleic acid in the presence of a polymerase, said first sequencing reagent comprising one or more nucleotide monomers, wherein said one or more nucleotide monomers pair with no more than three nucleotide types in said target, thereby forming a polynucleotide complementary to at least a portion of said target;
- and
- (b1) providing a second sequencing reagent to said target nucleic acid, said second sequencing reagent comprising at least one nucleotide monomer, said at least one nucleotide monomer of said second sequencing reagent comprising a reversibly terminating moiety, wherein said second sequencing reagent is provided subsequent to providing said first sequencing reagent, whereby sequence information for at least a portion of said target nucleic acid is obtained;
- or
- (b2) providing a second sequencing reagent to said target nucleic acid, said second sequencing reagent comprising at least one nucleotide monomer, wherein said at least one nucleotide monomer pairs with no more than three nucleotide types in said target, wherein said second sequencing reagent is provided subsequent to providing said first sequencing reagent, and wherein a signal that indicates the incorporation of said at least one nucleotide monomer into the polynucleotide is generated, whereby sequence information for at least a portion of said target nucleic acid is obtained.
5 more elements
A method of generating overlapping sequencing reads, said method comprising:
- providing a substrate having a surface, said surface comprising a first site and a second site, said first and second sites having a plurality of polynucleotides attached thereto and said first and second sites being spatially segregated from each other;
- and
- performing a sequencing run on said plurality of polynucleotides so as to produce an extended strand at said first and second spatially segregated sites, wherein the length of said extended strand produced at said first site is different from the length of said extended strand produced at said second site, thereby generating a set of overlapping sequencing reads, wherein performing a sequencing run comprises:
- (a) providing a first sequencing reagent to a target nucleic acid in the presence of a polymerase, said first sequencing reagent comprising one or more nucleotide monomers, wherein said one or more nucleotide monomers pair with no more than three nucleotide types in said target, thereby forming a polynucleotide complementary to at least a portion of said target;
- and
- (b1) providing a second sequencing reagent to said target nucleic acid, said second sequencing reagent comprising at least one nucleotide monomer, said at least one nucleotide monomer of said second sequencing reagent comprising a reversibly terminating moiety, wherein said second sequencing reagent is provided subsequent to providing said first sequencing reagent, whereby sequence information for at least a portion of said target nucleic acid is obtained;
- or
- (b2) providing a second sequencing reagent to said target nucleic acid, said second sequencing reagent comprising at least one nucleotide monomer, wherein said at least one nucleotide monomer pairs with no more than three nucleotide types in said target, wherein said second sequencing reagent is provided subsequent to providing said first sequencing reagent, and wherein a signal that indicates the incorporation of said at least one nucleotide monomer into the polynucleotide is generated, whereby sequence information for at least a portion of said target nucleic acid is obtained.
5 more elements
16-30. (canceled)
46-52. (canceled)
Dependencies are inferred from the claim text.