Sequence-Specific DNA Purification by Triplex Affinity Capture

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 89; no. 2; pp. 495 - 498
• Main Authors: Ito, Takashi, Smith, Cassandra L., Cantor, Charles R.
• Format: Journal Article
• Language: English
• Published: Washington, DC National Academy of Sciences of the United States of America 15.01.1992; National Acad Sciences; National Academy of Sciences
• Subjects: 550200 - Biochemistry; ANIMALS; AZOLES; Base Sequence; BASIC BIOLOGICAL SCIENCES; Biochemistry; Biological and medical sciences; BIOTIN; CARBOXYLIC ACIDS; CELL CONSTITUENTS; Chromatography, Affinity; CONCENTRATORS; CYTOLOGICAL TECHNIQUES; Deoxyribonucleic acid; Diverse techniques; DNA; DNA - chemistry; DNA - isolation & purification; DNA SEQUENCING; Fundamental and applied biological sciences. Psychology; Gels; Genomic Library; HETEROCYCLIC ACIDS; HETEROCYCLIC COMPOUNDS; Humans; Hydrogen-Ion Concentration; IMIDAZOLES; Libraries; MAGNETIC SEPARATORS; Magnetics; MAMMALS; MAN; Microsatellite repeats; Molecular and cellular biology; Molecular Sequence Data; Molecules; Nucleic Acid Conformation; NUCLEIC ACIDS; oligodeoxyribonucleotides; Oligodeoxyribonucleotides - chemistry; OLIGONUCLEOTIDES; ORGANIC ACIDS; ORGANIC COMPOUNDS; ORGANIC NITROGEN COMPOUNDS; ORGANIC SULFUR COMPOUNDS; PH VALUE; PLASMIDS; Polymerase Chain Reaction; PRIMATES; Repetitive Sequences, Nucleic Acid; STRUCTURAL CHEMICAL ANALYSIS; VERTEBRATES; VITAMIN B GROUP; VITAMINS; Molecular hybridization; Specificity; Magnetic separation; Nucleotide sequence; DNA; Triple helical structure; Affinity; Isolation; Oligonucleotide; Method
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.89.2.495

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)ndinucleotide repeats from a human genomic library. This procedure provides a prototype for other triplex-mediated DNA isolation technologies.