Methods and kits for nucleic acid sequencing

摘要

Various embodiments of the present disclosure generally relate to molecular biological protocols, equipment and reagents for the sequencing of target nucleic acid (DNA, RNA, cDNA, etc) molecules.

主权项

1. A method of sequencing a target polynucleotide, comprising:
immobilizing a primer along a nanowire, wherein the nanowire is coupled to a detector configured to detect a change in electrical resistance of the nanowire; hybridizing the target polynucleotide to the primer to form a primer-target polynucleotide immobilized along the nanowire; adding a nucleotide and a polymerase to the primer-target polynucleotide under conditions that support polymerization of a nascent chain when the added nucleotide is complementary to a base on the target polynucleotide downstream of the primer, wherein the added nucleotide comprises a reporter moiety and a linker, wherein the reporter moiety comprises an electrical charge that is sufficient to result in a detectable change in the resistance of the nanowire, and the reporter moiety is configured to protrude out from the nascent chain so as to reach-down toward the nanowire; and detecting, upon addition of said nucleotide, the change in the resistance of the nanowire, wherein the change in the resistance is characteristic of the nucleotide added to the nascent chain.