Patent ID: 6541227
Filing Date: 2003-04-01
Classification: C12P,C12Q

Abstract:
A method for preparing a labeled double-stranded DNA molecule by replacing a deoxyoligonucleotide sequence of a certain region of at least one of two single-stranded DNAs constituting a double-stranded DNA molecule with a deoxyoligonucleotide sequence having at least one labeled nucleotide, wherein the preparation method comprises the steps of:(A) incubating at least one type of deoxyoligonucleotide molecule having a sequence substantially equivalent to a certain region of one strand of the double-stranded DNA molecule and the double-stranded DNA molecule under a condition wherein said deoxyoligonucleotide molecule and double-stranded DNA molecule form a triple-stranded DNA at the 3â€² terminal region or the non-terminal region in a single-stranded DNA of the double-stranded DNA, and (B) incubating at least one type of complex of the double-stranded DNA and said at least one deoxyoligonucleotide having a region of the triple-stranded DNA formed in step (A) in the presence of four types of dNTPs comprising at least one labeled dNTP under a condition wherein the deoxyoligonucleotide sequence of the certain region of one strand of the double-stranded DNA molecule in said complex is substituted with a labeled deoxyoligonucleotide sequence, wherein said labeled deoxyoligonucleotide sequence has at least one labeled nucleotide, creating a labeled double-stranded DNA.