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Title: Triple helix purification and sequencing

Abstract

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:
Research Org.:
Univ. of Wisconsin, Madison, WI (United States)
OSTI Identifier:
869803
Patent Number(s):
5401632
Assignee:
Wisconsin Alumni Research Foundation (Madison, WI)
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:  
FG02-90ER61026
Resource Type:
Patent
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/

Citation Formats

Wang, Renfeng, Smith, Lloyd M, and Tong, Xinchun E. Triple helix purification and sequencing. United States: N. p., 1995. Web.
Wang, Renfeng, Smith, Lloyd M, & Tong, Xinchun E. Triple helix purification and sequencing. United States.
Wang, Renfeng, Smith, Lloyd M, and Tong, Xinchun E. Sun . "Triple helix purification and sequencing". United States. https://www.osti.gov/servlets/purl/869803.
@article{osti_869803,
title = {Triple helix purification and sequencing},
author = {Wang, Renfeng and Smith, Lloyd M and Tong, Xinchun E},
abstractNote = {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.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {1995},
month = {1}
}

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Works referenced in this record:

Spectroscopic and calorimetric investigation on the DNA triplex formed by d(CTCTTCTTTCTTTTCTTTCTTCTC) and d(GAGAAGAAAGA) at acidic pH
journal, January 1990


Molecular recognition by circular oligonucleotides. Strong binding of single-stranded DNA and RNA
journal, January 1991


Specificity and stringency in DNA triplex formation.
journal, November 1991


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


Molecular recognition by circular oligonucleotides: increasing the selectivity of DNA binding
journal, July 1991