Nucleic Acid Detection Methods
Abstract
The invention relates to methods for rapidly determining the sequence and/or length a target sequence. The target sequence may be a series of known or unknown repeat sequences which are hybridized to an array of probes. The hybridized array is digested with a single-strand nuclease and free 3'-hydroxyl groups extended with a nucleic acid polymerase. Nuclease cleaved heteroduplexes can be easily distinguish from nuclease uncleaved heteroduplexes by differential labeling. Probes and target can be differentially labeled with detectable labels. Matched target can be detected by cleaving resulting loops from the hybridized target and creating free 3-hydroxyl groups. These groups are recognized and extended by polymerases added into the reaction system which also adds or releases one label into solution. Analysis of the resulting products using either solid phase or solution. These methods can be used to detect characteristic nucleic acid sequences, to determine target sequence and to screen for genetic defects and disorders. Assays can be conducted on solid surfaces allowing for multiple reactions to be conducted in parallel and, if desired, automated.
- Inventors:
-
- Boston, MA
- Brookline, MA
- Issue Date:
- Research Org.:
- Boston University
- Sponsoring Org.:
- USDOE
- OSTI Identifier:
- 879271
- Patent Number(s):
- 5753439
- Application Number:
- 08/446102
- Assignee:
- Trustees of Boston University (Boston, MA)
- Patent Classifications (CPCs):
-
C - CHEMISTRY C12 - BIOCHEMISTRY C12Q - MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
- DOE Contract Number:
- FG02-93ER61609
- Resource Type:
- Patent
- Country of Publication:
- United States
- Language:
- English
Citation Formats
Smith, Cassandra L, Yaar, Ron, Szafranski, Przemyslaw, and Cantor, Charles R. Nucleic Acid Detection Methods. United States: N. p., 1998.
Web.
Smith, Cassandra L, Yaar, Ron, Szafranski, Przemyslaw, & Cantor, Charles R. Nucleic Acid Detection Methods. United States.
Smith, Cassandra L, Yaar, Ron, Szafranski, Przemyslaw, and Cantor, Charles R. Tue .
"Nucleic Acid Detection Methods". United States. https://www.osti.gov/servlets/purl/879271.
@article{osti_879271,
title = {Nucleic Acid Detection Methods},
author = {Smith, Cassandra L and Yaar, Ron and Szafranski, Przemyslaw and Cantor, Charles R},
abstractNote = {The invention relates to methods for rapidly determining the sequence and/or length a target sequence. The target sequence may be a series of known or unknown repeat sequences which are hybridized to an array of probes. The hybridized array is digested with a single-strand nuclease and free 3'-hydroxyl groups extended with a nucleic acid polymerase. Nuclease cleaved heteroduplexes can be easily distinguish from nuclease uncleaved heteroduplexes by differential labeling. Probes and target can be differentially labeled with detectable labels. Matched target can be detected by cleaving resulting loops from the hybridized target and creating free 3-hydroxyl groups. These groups are recognized and extended by polymerases added into the reaction system which also adds or releases one label into solution. Analysis of the resulting products using either solid phase or solution. These methods can be used to detect characteristic nucleic acid sequences, to determine target sequence and to screen for genetic defects and disorders. Assays can be conducted on solid surfaces allowing for multiple reactions to be conducted in parallel and, if desired, automated.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {Tue May 19 00:00:00 EDT 1998},
month = {Tue May 19 00:00:00 EDT 1998}
}
Works referenced in this record:
The clinical chemist
journal, December 1994
- Burns, David E.
- Clinical Chemistry, Vol. 40, Issue 12, p. 2332-2334
Dynamic mutations: A new class of mutations causing human disease
journal, September 1992
- Richards, Robert I.; Sutherland, Grant R.
- Cell, Vol. 70, Issue 5
Simple tandem DNA repeats and human genetic disease.
journal, April 1995
- Sutherland, G. R.; Richards, R. I.
- Proceedings of the National Academy of Sciences, Vol. 92, Issue 9
Advances in genosensor research
journal, May 1995
- Beattie, K. L.; Beattie, W. G.; Meng, L.
- Clinical Chemistry, Vol. 41, Issue 5
Oligonucleotide Arrays: New Concepts and Possibilities
journal, November 1994
- Chetverin, Alexander B.; Kramer, Fred Russell
- Bio/Technology, Vol. 12, Issue 11
Triplet repeat mutations in human disease
journal, May 1992
- Caskey, C.; Pizzuti, A.; Fu, Y.
- Science, Vol. 256, Issue 5058
Trinucleotide repeat instability: when and where?
journal, June 1993
- Nelson, David L.; Warren, Stephen T.
- Nature Genetics, Vol. 4, Issue 2
Two-dimensional S1 nuclease heteroduplex mapping: detection of rearrangements in bacterial genomes.
journal, May 1984
- Yee, T.; Inouye, M.
- Proceedings of the National Academy of Sciences, Vol. 81, Issue 9
A rapid scanning strip for tri- and dinucleotide short tandem repeats
journal, January 1994
- Wehnert, Manfred S.; Matson, Robert S.; Rampal, Jang B.
- Nucleic Acids Research, Vol. 22, Issue 9
Genes with triplet repeats: candidate mediators of neuropsychiatric disorders
journal, July 1993
- Ross, Christopher A.; McInnis, Melvin G.; Margolis, Russell L.
- Trends in Neurosciences, Vol. 16, Issue 7