Rapid purification of circular DNA by triplex-mediated affinity capture
Abstract
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:
-
- 4817 Sheboygan Ave., Madison, WI 53705
- 1115 Amherst Dr., Madison, WI 53705
- Issue Date:
- Research Org.:
- Univ. of Wisconsin, Madison, WI (United States)
- OSTI Identifier:
- 870773
- Patent Number(s):
- 5591841
- Assignee:
- Ji, Huamin (4817 Sheboygan Ave., Madison, WI 53705);Smith, Lloyd M. (1115 Amherst Dr., Madison, WI 53705)
- Patent Classifications (CPCs):
-
C - CHEMISTRY C12 - BIOCHEMISTRY C12N - MICROORGANISMS OR ENZYMES
C - CHEMISTRY C12 - BIOCHEMISTRY C12Q - MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
- DOE Contract Number:
- FG02-90ER61026
- Resource Type:
- Patent
- 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/
Citation Formats
Ji, Huamin, and Smith, Lloyd M. Rapid purification of circular DNA by triplex-mediated affinity capture. United States: N. p., 1997.
Web.
Ji, Huamin, & Smith, Lloyd M. Rapid purification of circular DNA by triplex-mediated affinity capture. United States.
Ji, Huamin, and Smith, Lloyd M. Wed .
"Rapid purification of circular DNA by triplex-mediated affinity capture". United States. https://www.osti.gov/servlets/purl/870773.
@article{osti_870773,
title = {Rapid purification of circular DNA by triplex-mediated affinity capture},
author = {Ji, Huamin and Smith, Lloyd M},
abstractNote = {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.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {1997},
month = {1}
}
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