Field switch sequencing

摘要

The present invention provides novel compositions, methods and apparatus for DNA sequencing that can be performed, e.g., in a two-electrode chamber. The present invention also provides a method for sequencing a nucleic acid comprising immobilizing a plurality of complexes comprising a target nucleic acid, a primer nucleic acid, and a polymerase onto a surface, contacting the surface with a plurality of charged particles comprising a nucleotide phosphate by applying an electric field, reversing the electric field to transport unbound charged particles away from the surface, and detecting the incorporation of a nucleotide phosphate into a single molecule of the primer nucleic acid.

主权项

1. A method for increasing the signal to noise ratio (S/N) for single particle detection while sequencing a nucleic acid, said method comprising:
(a) immobilizing a plurality of complexes comprising a target nucleic acid, a primer nucleic acid, and a polymerase onto a surface; (b) providing a sequencing reaction mixture including single particles, which are dendrimers, each said single particle comprising both i) a plurality of dyes and ii) a plurality of incorporatable substrate nucleotide phosphates for the polymerase, wherein a plurality of said single particles sequentially reacts with an immobilized complex, wherein in each said sequential reaction one of the plurality of nucleotide phosphates reacts with the polymerase, is cleaved from the dendrimer, and is incorporated into the primer nucleic acid, thereby releasing the dendrimer from the complex; and (c) detecting said single particle while incorporating each nucleotide phosphate, thereby sequencing the target nucleic acid.