Specific terminal labeling of DNA molecules.
We report a simple method to directly label or modify a specific terminus of linear DNA molecules. The method is based upon our finding that a presumably triple-stranded structure by RecA-mediated formation at the terminus formed with deoxyoligonucleotides, whose sequence is complementary to the 5' terminus of one of the strands of a double-stranded DNA molecule, is quite stable and can serve as a template for DNA polymerase reaction, with the nucleotides being incorporated by an exchange reaction. This novel type of nucleotide incorporation has made it possible to label a specific terminus of target double-stranded DNA molecules by a direct means (without amplification) regardless of its molecular size, a procedure previously unavailable. As an application, we show that large DNA molecules can be fixed to a solid support in a specific orientation, thus being utilized for various analytical purposes of DNA molecules.
Shigemori
Yasushi
Y
Aisin Cosmos R&D Co. Ltd., Chiba, Chiba, Japan.
Oishi
Michio
M
D001483:Base Sequence; D004279:DNA, Viral / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism*; D009711:Nucleotides / Q000378:metabolism; D013194:Staining and Labeling / Q000379:methods*; D013698:Templates, Genetic
D004279:DNA, Viral; D009711:Nucleotides