Specific terminal labeling of DNA molecules.

BioTechniques (2007), Volume 42, Page 622

Abstract:

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.

Polymerases:

Topics:

Nucleotide Incorporation, Nucleotide Analogs / Template Lesions, Exonuclease Activity

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