NUCLEIC ACID SEQUENCING USING NICKING AGENTS

摘要

Nucleotide base sequence information is obtained upon combining a template nucleic acid comprising a primer strand and a target nucleic acid strand, the target nucleic acid strand comprising an interrogation sequence, with a nicking agent that recognizes a double-stranded sequence present in the template nucleic acid and then nicks the primer strand of the template nucleic acid at a location relative to the nicking agent recognition sequence, with a distributive polymerase that will extend the primer strand from the site where the nicking agent nicked the primer strand, in the presence of suitably-reactive mononucleotides, to provide a plurality of oligonucleotides that are complements of the sequence present in the interrogation sequence, where the plurality differ in having different nucleotide lengths, so the aligning the plurality of oligonucleotides provides nucleotide base sequence information about the interrogation sequence.