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Title: DNA purification by triplex-affinity capture and affinity capture electrophoresis

Abstract

The invention provides a method for purifying or isolating double stranded DNA intact using triple helix formation. The method includes the steps of complexing an oligonucleotide and double stranded DNA to generate a triple helix and immobilization of the triple helix on a solid phase by means of a molecular recognition system such as avidin/biotin. The purified DNA is then recovered intact by treating the solid phase with a reagent that breaks the bonds between the oligonucleotide and the intact double stranded DNA while not affecting the Watson-Crick base pairs of the double helix. The present invention also provides a method for purifying or isolating double stranded DNA intact by complexing the double stranded DNA with a specific binding partner and recovering the complex during electrophoresis by immobilizing it on a solid phase trap imbedded in an electrophoretic gel.

Inventors:
 [1];  [2];  [1]
  1. Boston, MA
  2. Chiba, JP
Issue Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
OSTI Identifier:
870243
Patent Number(s):
5482836
Assignee:
Regents of University of California (Oakland, CA)
Patent Classifications (CPCs):
C - CHEMISTRY C12 - BIOCHEMISTRY C12Q - MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
C - CHEMISTRY C12 - BIOCHEMISTRY C12N - MICROORGANISMS OR ENZYMES
DOE Contract Number:  
AC03-76SF00098
Resource Type:
Patent
Country of Publication:
United States
Language:
English
Subject:
dna; purification; triplex-affinity; capture; affinity; electrophoresis; provides; method; purifying; isolating; double; stranded; intact; triple; helix; formation; steps; complexing; oligonucleotide; generate; immobilization; solid; phase; means; molecular; recognition; avidin; biotin; purified; recovered; treating; reagent; breaks; bonds; affecting; watson-crick; base; pairs; specific; binding; partner; recovering; complex; immobilizing; trap; imbedded; electrophoretic; triple helix; stranded dna; binding partner; solid phase; double stranded; affinity capture; double helix; base pairs; base pair; specific binding; /435/204/999/

Citation Formats

Cantor, Charles R, Ito, Takashi, and Smith, Cassandra L. DNA purification by triplex-affinity capture and affinity capture electrophoresis. United States: N. p., 1996. Web.
Cantor, Charles R, Ito, Takashi, & Smith, Cassandra L. DNA purification by triplex-affinity capture and affinity capture electrophoresis. United States.
Cantor, Charles R, Ito, Takashi, and Smith, Cassandra L. Mon . "DNA purification by triplex-affinity capture and affinity capture electrophoresis". United States. https://www.osti.gov/servlets/purl/870243.
@article{osti_870243,
title = {DNA purification by triplex-affinity capture and affinity capture electrophoresis},
author = {Cantor, Charles R and Ito, Takashi and Smith, Cassandra L},
abstractNote = {The invention provides a method for purifying or isolating double stranded DNA intact using triple helix formation. The method includes the steps of complexing an oligonucleotide and double stranded DNA to generate a triple helix and immobilization of the triple helix on a solid phase by means of a molecular recognition system such as avidin/biotin. The purified DNA is then recovered intact by treating the solid phase with a reagent that breaks the bonds between the oligonucleotide and the intact double stranded DNA while not affecting the Watson-Crick base pairs of the double helix. The present invention also provides a method for purifying or isolating double stranded DNA intact by complexing the double stranded DNA with a specific binding partner and recovering the complex during electrophoresis by immobilizing it on a solid phase trap imbedded in an electrophoretic gel.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {1996},
month = {1}
}

Works referenced in this record:

Magnesium ion-dependent triple-helix structure formed by homopurine-homopyrimidine sequences in supercoiled plasmid DNA.
journal, June 1988


Cleaving yeast and Escherichia coli genomes at a single site
journal, October 1990


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


Structure, stability, and thermodynamics of a short intermolecular purine-purine-pyrimidine triple helix
journal, June 1991


Preparative separation of the complementary strands of DNA restriction fragments by alkaline RPC-5 chromatography
journal, January 1978


Single-site enzymatic cleavage of yeast genomic DNA mediated by triple helix formation
journal, March 1991


Selective cleavage of human DNA: RecA-assisted restriction endonuclease (RARE) cleavage
journal, December 1991


A combinatorial approach toward DNA recognition
journal, September 1991


Mechanism of integrating foreign DNA during transformation of Bacillus subtilis.
journal, August 1978


Recognition of mixed-sequence duplex DNA by alternate-strand triple-helix formation
journal, March 1990


Separation of random fragments of DNA according to properties of their sequences.
journal, August 1980


Formation of intramolecular triplex in homopurine-homopyrimidine mirror repeats with point substitutions
journal, January 1990


Binding of triple helix forming oligonucleotides to sites in gene promoters
journal, September 1991


Sequence-specific cleavage of double helical DNA by triple helix formation
journal, October 1987


The chemistry and biology of unusual DNA structures adopted by oligopurine · oligopyrimidine sequences
journal, November 1988


A structural basis for S1 nuclease sensitivity of double-stranded DNA
journal, August 1985


Selective enrichment of a large size genomic DNA fragment by affinity capture: an approach for genome mapping
journal, January 1990


Oligonucleotide inhibition of IL2Rα mRNA transcription by promoter region collinear triplex formation in lymphocytes
journal, January 1991


Site-specific oligonucleotide binding represses transcription of the human c-myc gene in vitro
journal, July 1988