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Nucleic acid detection methods

United States Patent

5,753,439
May 19, 1998
View the Complete Patent at the US Patent & Trademark Office
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.
Smith; Cassandra L. (Boston, MA), Yaar; Ron (Brookline, MA), Szafranski; Przemyslaw (Boston, MA), Cantor; Charles R. (Boston, MA)
Trustees of Boston University (Boston, MA)
08/ 446,102
May 19, 1995
RIGHTS TO THE INVENTION This invention was made with United States Government support under grant number DE-FG02-93ER61609, awarded by the United States Department of Energy, and grant number AIBS2154, awarded by the United States Department of the Army, and the United States Government has certain rights in the invention.