Patent ID: 8592183

Claim:
A method for sequencing a target polynucleotide, comprising: a. immobilizing the target polynucleotide on a support of an apparatus for sequencing a polynucleotide, comprising: i. a flow cell comprising a fluid flow path; ii. a monochromatic laser; and iii. a total internal reflection microscope, wherein the support is positioned within the fluid flow path of the flow cell such that, upon immobilization of a target polynucleotide, the target polynucleotide is positioned in a reaction site within at least a portion of the fluid flow path; and wherein the laser is configured to emit an optical beam directed at the reaction site, and wherein a total internal reflection microscope is focused on the reaction site such that a polymerization reaction associated with the target polynucleotide can be monitored over time; b. contacting the target polynucleotide with a polymerase enzyme and a nucleotide selected from the group consisting of A, T(U), G, and C under conditions suitable for a polymerase reaction to proceed, wherein the nucleotide comprises a removable blocking group; c. observing the interaction of the polymerase enzyme and the nucleotide with the target polynucleotide; d. determining whether the polymerase has incorporated the nucleotide onto the primer sequence; and e. repeating steps (b)-(d) one or more times, thereby identifying the sequence of the target polynucleotide.