US 9,005,929 B2 · Multibase delivery for long reads in sequencing by synthesis protocols

Abstract

A method for obtaining nucleic acid sequence information that can include steps of (a) providing a first sequencing reagent to a target nucleic acid, wherein the first sequencing reagent comprises at least two different nucleotide monomers, (b) detecting the incorporation of a nucleotide monomer present in the first sequencing reagent into a polynucleotide strand complementary to at least a portion of the target nucleic acid, (c) providing a second sequencing reagent to said target nucleic acid, wherein the second sequencing reagent comprises one or more nucleotide monomers, at least one of the one or more nucleotide monomers being different from the nucleotide monomers present in the first sequencing reagent, and wherein the second sequencing reagent is provided subsequent to providing the first sequencing reagent, and (d) detecting the incorporation of a nucleotide monomer present in the second sequencing reagent into the polynucleotide strand.

Claims32

Dependent claims

A method for obtaining nucleic acid sequence information, said method comprising the steps of:

(a) providing a first sequencing reagent to a target nucleic acid, wherein said first sequencing reagent comprises at least two different nucleotide monomers;
(b) detecting the incorporation of a nucleotide monomer present in said first sequencing reagent into a polynucleotide strand complementary to at least a portion of said target nucleic acid;
(c) providing a second sequencing reagent to said target nucleic acid, wherein said second sequencing reagent comprises one or more nucleotide monomers, at least one of said one or more nucleotide monomers being different from the nucleotide monomers present in said first sequencing reagent, and wherein said second sequencing reagent is provided subsequent to providing said first sequencing reagent;
(d) detecting the incorporation of a nucleotide monomer present in said second sequencing reagent into the polynucleotide strand;
and
(e) repeating steps (a) through (d) for said target nucleic acid, thereby obtaining sequence information for at least a portion of said target nucleic acid.

3 more elements

A method for obtaining nucleic acid sequence information, said method comprising the steps of:

(a) detecting the incorporation of a nucleotide monomer present in a first sequencing reagent into a first polynucleotide strand complementary to at least a portion of a target nucleic acid, wherein said first sequencing reagent comprises at least two different nucleotide monomers;
(b) detecting the incorporation of a nucleotide monomer present in a second sequencing reagent into the first polynucleotide strand, wherein said second sequencing reagent comprises one or more nucleotide monomers, at least one of said one or more nucleotide monomers being different from the nucleotide monomers present in said first sequencing reagent;
(c) removing said first polynucleotide strand;
and
(d) detecting the incorporation of a nucleotide monomer present in a third sequencing reagent into a second polynucleotide strand complementary to at least a portion of said target nucleic acid, wherein said third sequencing reagent comprises two or more nucleotide monomers, wherein at least one of said two or more nucleotide monomers is different from the at least two different nucleotide monomers present in said first sequencing reagent and at least one of said two or more nucleotide monomers is different from the one or more different nucleotide monomers present in said second sequencing reagent;
and
(e) repeating steps (a) through (d) for said target nucleic acid.

4 more elements

Dependencies are inferred from the claim text.

Multibase delivery for long reads in sequencing by synthesis protocols

Life of this patent

Multibase delivery for long reads in sequencing by synthesis protocols

Life of this patent

Abstract

Claims32