Sequence-specific DNA purification by triplex affinity capture.

T Ito; C L Smith; C R Cantor

doi:10.1073/pnas.89.2.495

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Sequence-specific DNA purification by triplex affinity capture.

T Ito·C L Smith·C R Cantor

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Abstract

A DNA isolation procedure was developed by using triple-helix formation and magnetic separation. In this procedure, target DNA is captured by a biotinylated oligonucleotide via intermolecular triplex formation, bound to streptavidin-coated magnetic beads, and recovered in double-stranded form by elution with a mild alkaline buffer that destabilizes the triple helix. The effectiveness of the procedure was demonstrated by a model experiment with an artificially reconstructed library and, also, by the isolation of (dT-dC)n.(dG-dA)n dinucleotide repeats from a human genomic library. This procedure provides a prototype for other triplex-mediated DNA isolation technologies.

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Base Sequence Biotin Chromatography, Affinity DNA Genomic Library Humans Hydrogen-Ion Concentration Magnetics Molecular Sequence Data Nucleic Acid Conformation Oligodeoxyribonucleotides Polymerase Chain Reaction Repetitive Sequences, Nucleic Acid Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.Research Support, U.S. Gov't, P.H.S.

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T Ito, C L Smith, C R Cantor. 1992-01-15. Sequence-specific DNA purification by triplex affinity capture.. https://doi.org/10.1073/pnas.89.2.495

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