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

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:
OSTI Identifier:
870243
Assignee:
Regents of University of California (Oakland, CA) LBNL
Patent Number(s):
US 5482836
Contract Number:
AC03-76SF00098
Research Org:
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA
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/