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Title: 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:
 [1];  [2]
  1. 4817 Sheboygan Ave., Madison, WI 53705
  2. 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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Works referenced in this record:

Rapid purification of double-stranded DNA by triple-helix-mediated affinity capture
journal, May 1993


Covalent Attachment of DNA to Agarose. Improved Synthesis and Use in Affinity Chromatography
journal, June 1975


A stable complex between homopyrimidine oligomers and the homologous regions of duplex DNAs
journal, January 1988


Affinity capture electrophoresis for sequence-specific DNA purification
journal, June 1992


Rapid isolation of cosmid insert DNA by triple-helix-mediated affinity capture
journal, January 1994


Sequence-specific DNA purification by triplex affinity capture.
journal, January 1992


Triplex affinity capture of a single copy clone from a yeast genomic library
journal, January 1992