Sequence-specific DNA purification by triplex affinity capture.
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.
Ito
T
T
Department of Molecular and Cell Biology, University of California, Berkeley 94720.
Smith
C L
CL
Cantor
C R
CR
D001483:Base Sequence; D001710:Biotin; D002846:Chromatography, Affinity; D004247:DNA / Q000737:chemistry / Q000302:isolation & purification*; D015698:Genomic Library; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008280:Magnetics; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D016133:Polymerase Chain Reaction; D012091:Repetitive Sequences, Nucleic Acid
D009838:Oligodeoxyribonucleotides; D001710:Biotin; D004247:DNA