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Triple helix purification and sequencing

Patent ·
OSTI ID:869803
 [1];  [2];  [2]
  1. Dublin, CA
  2. Madison, WI
+ Show Author Affiliations

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.

Research Organization:
University of Wisconsin
DOE Contract Number:
FG02-90ER61026
Assignee:
Wisconsin Alumni Research Foundation (Madison, WI)
Patent Number(s):
US 5401632
OSTI ID:
869803
Country of Publication:
United States
Language:
English

References (8)

Molecular recognition by circular oligonucleotides: increasing the selectivity of DNA binding journal July 1991
NMR structural studies of intramolecular (Y+)n.cntdot.(R+)n(Y-)n DNA triplexes in solution: imino and amino proton and nitrogen markers of G.cntdot.AT base triple formation journal September 1991
Intramolecular triplex formation of the purine-purine-pyrimidine type journal May 1991
Magnetic DNA hybridization properties of oligonucleotide probes attached to superparamagnetic beads and their use in the isolation of poly(A) mRNA from eukaryotic cells journal October 1990
Sequence-specific DNA purification by triplex affinity capture. journal January 1992
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
Spectroscopic and calorimetric investigation on the DNA triplex formed by d(CTCTTCTTTCTTTTCTTTCTTCTC) and d(GAGAAGAAAGA) at acidic pH journal January 1990

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Related Subjects

/435/999/
anchor
bound
circular
circular dna
complementary
complex
desired
disclosed
dna
dna sequencing
dna synthesis
double
double helix
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helix
hybridization
hybridizes
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magnet
magnetic
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oligo
oligonucleotide
primer
provided
purification
purify
purifying
purposes
region
regions
sequencing
single
single strand
single stranded
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stable
stranded
synthesis
template
templates
triple
triple helix