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Updated: Jul 14, 2026
12:07
Sequence-specific Labeling of Nucleic Acids and Proteins with Methyltransferases and Cofactor Analogues
Published on: November 22, 2014
Specific terminal labeling of DNA molecules
Yasushi Shigemori1, Michio Oishi
1Aisin Cosmos R&D Co. Ltd., Chiba, Chiba, Japan.
Biotechniques
|May 23, 2007
Summary
Researchers developed a novel method to directly label DNA termini using RecA protein and a stable triple-stranded structure. This technique enables specific nucleotide incorporation for DNA modification and orientation on solid supports.
Area of Science:
- Molecular Biology
- Biochemistry
- Genetics
Background:
- Directly labeling or modifying specific DNA termini is challenging.
- Existing methods often require amplification or are size-limited.
Purpose of the Study:
- To develop a simple, direct method for labeling or modifying specific termini of linear DNA molecules.
- To enable specific nucleotide incorporation into DNA termini regardless of molecular size.
Main Methods:
- Utilized RecA-mediated formation of a stable triple-stranded structure at the DNA terminus using complementary deoxyoligonucleotides.
- Employed this structure as a template for DNA polymerase-mediated nucleotide exchange reactions.
Main Results:
- Successfully demonstrated direct labeling of specific DNA termini without amplification.
- Showcased the ability to orient large DNA molecules on solid supports for analytical applications.
Conclusions:
- The developed method offers a novel and versatile approach for specific DNA terminus modification.
- This technique has significant potential for various DNA analysis and manipulation applications.
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