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Title: Rapid purification of circular DNA by triplex-mediated affinity capture

A single-step capture of a target supercoiled double-stranded DNA molecule is accomplished by forming a local triple-helix among two strands of the supercoiled circular DNA and an oligonucleotide probe. The oligonucleotide is bound to an immobilizing support which facilitates the immobilization and purification of target DNA molecules. Non-target DNA molecules and other contaminating cellular material are easily removed by washing. The triple-helical structure is destabilized by raising the pH, leaving purified target DNA in the supernatant and reusable affinity capture oligonucleotide secured to the immobilizing support.
Inventors:
 [1];  [2]
  1. (4817 Sheboygan Ave., Madison, WI 53705)
  2. (1115 Amherst Dr., Madison, WI 53705)
Issue Date:
OSTI Identifier:
870773
Assignee:
Ji, Huamin (4817 Sheboygan Ave., Madison, WI 53705);Smith, Lloyd M. (1115 Amherst Dr., Madison, WI 53705) CHO
Patent Number(s):
US 5591841
Contract Number:
FG02-90ER61026
Research Org:
University of Wisconsin
Country of Publication:
United States
Language:
English
Subject:
rapid; purification; circular; dna; triplex-mediated; affinity; capture; single-step; target; supercoiled; double-stranded; molecule; accomplished; forming; local; triple-helix; strands; oligonucleotide; probe; bound; immobilizing; support; facilitates; immobilization; molecules; non-target; contaminating; cellular; material; easily; removed; washing; triple-helical; structure; destabilized; raising; ph; leaving; purified; supernatant; reusable; secured; cellular material; stranded dna; double-stranded dna; easily removed; dna molecules; dna molecule; target dna; circular dna; affinity capture; oligonucleotide probe; /999/435/