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Title: 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{prime}-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. 18 figs.

Inventors:
; ; ;
Issue Date:
Research Org.:
Boston University
Sponsoring Org.:
USDOE, Washington, DC (United States); Department of the Army, Washington, DC (United States)
OSTI Identifier:
644363
Patent Number(s):
5753439
Application Number:
PAN: 8-446,102; CNN: Grant AIBS2154
Assignee:
Boston Univ., MA (United States)
DOE Contract Number:  
FG02-93ER61609
Resource Type:
Patent
Resource Relation:
Other Information: PBD: 19 May 1998
Country of Publication:
United States
Language:
English
Subject:
55 BIOLOGY AND MEDICINE, BASIC STUDIES; DNA SEQUENCING; DNA HYBRIDIZATION; NUCLEASES; POLYMERASES; AMINO ACID SEQUENCE

Citation Formats

Smith, C L, Yaar, R, Szafranski, P, and Cantor, C R. Nucleic acid detection methods. United States: N. p., 1998. Web.
Smith, C L, Yaar, R, Szafranski, P, & Cantor, C R. Nucleic acid detection methods. United States.
Smith, C L, Yaar, R, Szafranski, P, and Cantor, C R. Tue . "Nucleic acid detection methods". United States.
@article{osti_644363,
title = {Nucleic acid detection methods},
author = {Smith, C L and Yaar, R and Szafranski, P and Cantor, C 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{prime}-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. 18 figs.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {1998},
month = {5}
}