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US 6,541,226 B1Granted
Method for specifically cleaving double-stranded DNA and kit for the method
AssigneeAisin Seiki Kabushiki Kaisha
InventorsYasushi Shigemori, Michio Oishi
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Abstract
A method for specifically cleaving a double-stranded DNA. The method comprises forming a three-stranded portion comprising the double-stranded DNA portion including the position to be cleaved or its vicinity and an oligonucleotide and treating the three-stranded protion with a nuclease.
Claims11
A method for specifically cleaving a double-stranded DNA using an enzyme, comprising the steps of:
- (A) forming a complex having a three-stranded nucleic acid portion with a double-stranded DNA to be cleaved in the presence of a homologous recombinant protein, a nucleoside triphosphate or analogs thereof, wherein the three-stranded nucleic acid portion comprises a double-stranded DNA and a single-stranded DNA or PNA molecule that are 100% complementary to each other and are 30 or more nucleotides in length;
- (B) cleaving the complex thus obtained with a nuclease that recognizes the three-stranded nucleic acid portion in said complex and cleaves a phosphodiester bond of the double-stranded DNA in or adjacent to the three-stranded nucleic portion;
- and
- (C) inactivating the nuclease.
1 more element
Dependencies are inferred from the claim text.