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Title: Triple helix purification and sequencing

Disclosed herein are methods, kits, and equipment for purifying single stranded circular DNA and then using the DNA for DNA sequencing purposes. Templates are provided with an insert having a hybridization region. An elongated oligonucleotide has two regions that are complementary to the insert and the oligo is bound to a magnetic anchor. The oligo hybridizes to the insert on two sides to form a stable triple helix complex. The anchor can then be used to drag the template out of solution using a magnet. The system can purify sequencing templates, and if desired the triple helix complex can be opened up to a double helix so that the oligonucleotide will act as a primer for further DNA synthesis.
Inventors:
 [1];  [2];  [2]
  1. (Dublin, CA)
  2. (Madison, WI)
Issue Date:
OSTI Identifier:
869803
Assignee:
Wisconsin Alumni Research Foundation (Madison, WI) CHO
Patent Number(s):
US 5401632
Contract Number:
FG02-90ER61026
Research Org:
University of Wisconsin
Country of Publication:
United States
Language:
English
Subject:
triple; helix; purification; sequencing; disclosed; methods; kits; equipment; purifying; single; stranded; circular; dna; purposes; templates; provided; insert; hybridization; region; elongated; oligonucleotide; regions; complementary; oligo; bound; magnetic; anchor; hybridizes; form; stable; complex; drag; template; solution; magnet; purify; desired; double; primer; synthesis; triple helix; single stranded; dna synthesis; dna sequencing; circular dna; double helix; single strand; /435/999/