Compositions, systems, and methods for sequencing polynucleotides using tethers anchored to polymerases adjacent to nanopores
A composition includes a nanopore including a first side, a second side, and an aperture extending through the first and second sides. The composition includes a plurality of nucleotides, wherein each of the nucleotides comprises an elongated tag that corresponds to the type of nucleotide, the elong...
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| Format: | Patent |
| Sprache: | eng ; heb |
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| Zusammenfassung: | A composition includes a nanopore including a first side, a second side, and an aperture extending through the first and second sides. The composition includes a plurality of nucleotides, wherein each of the nucleotides comprises an elongated tag that corresponds to the type of nucleotide, the elongated tag being sufficiently long and narrow to be disposed within at least a portion of the aperture of the nanopore. The composition includes first and second polynucleotides, the first polynucleotide being complementary to the second polynucleotide. The composition includes a polymerase disposed adjacent to the first side of the nanopore, the polymerase configured to add nucleotides of the plurality of nucleotides to the first polynucleotide based on a sequence of the second polynucleotide. The composition includes a permanent tether including a head region, a tail region, and an elongated body disposed therebetween, the head region being anchored to the polymerase, wherein the elongated body occurs in the aperture of the nanopore. The composition includes a first moiety disposed on the elongated body, wherein the first moiety is configured to bind to the elongated tag of a first nucleotide upon which the polymerase is acting, the first moiety being configured to generate a first signal state responsive to the polymerase acting upon the first nucleotide, the first nucleotide being identifiable based on the first signal state. The composition includes a reporter region disposed on the elongated body; wherein the reporter region is configured to generate a second signal state responsive to release of pyrophosphate responsive to the polymerase successfully incorporating the first nucleotide into the first polynucleotide, it being detectable based upon the second signal state whether the first nucleotide is complementary or is not complementary to a next nucleotide in the sequence of the second polynucleotide. |
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