Patent ID: 6121001
Filing Date: 2000-09-19
Classification: C12Q

Abstract:
A method for modifying an oligonucleotide, the method comprising incubating under isothermal conditions the oligonucleotide, a polynucleotide, and a nuclease, wherein the oligonucleotide and the polynucleotide form a complex comprising said oligonucleotide and said polynucleotide in which at least a portion of the oligonucleotide is hybridized to the polynucleotide, wherein the isothermal conditions are at or near the melting temperature of the complex and wherein the oligonucleotide, when the portion is hybridized to the polynucleotide, is cleaved by the nuclease to provide (i) a first fragment that is substantially non-hybridizable to the polynucleotide and includes no more than five nucleotides from the 5'-end of the portion and (ii) a second fragment that is 3' of the first fragment with reference to the intact oligonucleotide and is substantially hybridizable to the polynucleotide, thereby modifying said oligonucleotide, wherein said first fragment and said second fragment are continuously produced under said isothermal conditions.