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US 6,297,010 B1Granted
Method for detecting and identifying mutations
AssigneeGenzyme Corporation
InventorJames E. Stefano
Life of this patent
Estimating the term…
Abstract
This invention provides various methods for identifying one or more genetic alterations in a sample polynucleotide strand.
Claims21
A method for identifying one or more genetic alterations in a sample polynucleotide strand, comprising:
- (a) providing a duplex comprising the sample polynucleotide strand and a reference polynucleotide strand;
- (b) contacting the duplex with an agent which recognizes and protects base pair mismatches under conditions which allow the agent to bind to and protect the duplex at the mismatch to form a duplex:agent complex;
- (c) removing unprotected base pairs;
- (d) providing a preselected site from which to sequence;
- and
- (e) determining the sequence of the sample strand to identify the one or more genetic alterations in the sample polynucleotide strand.
3 more elements
A method for identifying one or more genetic alterations in a sample polynucleotide strand, comprising:
- (a) providing a plurality of duplexes, wherein each duplex comprises a sample polynucleotide strand and a reference polynucleotide strand;
- (b) contacting the duplexes with an agent which recognizes and protects base pair mismatches under conditions which allow the agent to bind to and protect the duplexes at the mismatch to form duplex:agent complexes;
- (c) removing unprotected base pairs;
- (d) providing a preselected site from which to sequence;
- and
- (e) determining the sequence of the sample strands to identify the one or more genetic alterations in the sample polynucleotide strands.
3 more elements
A method for identifying one or more genetic alteration(s) in a sample nucleotide strand comprising:
- (a) providing a duplex by immobilizing the sample polynucleotide strand on one or more solid supports and contacting the sample polynucleotide strand with a reference polynucleotide strand under conditions suitable to form a duplex between the sample and reference strands;
- (b) contacting the duplex with an agent which recognizes and protects base pair mismatches under conditions which allow the agent to bind to and protect the duplex at the mismatch to form a duplex:agent complex;
- (c) removing unprotected base pairs;
- (d) providing a preselected site from which to sequence;
- and
- (e) sequencing the sample polynucleotide strands to identify the one or more genetic alteration(s).
3 more elements
A method for identifying one or more genetic alteration(s) in one or more sample polynucleotide strands comprising:
- (a) providing a plurality of duplexes by immobilizing a plurality of sample polynucleotide strands on one or more solid supports and contacting the plurality of sample polynucleotide strands with a reference polynucleotide strand under conditions suitable to form a duplex between the sample and reference strands;
- (b) contacting the duplex with an agent which recognizes and protects base pair mismatches under conditions which allow the agent to bind to and protect the duplex at the mismatch to form a duplex:agent complex;
- (c) removing unprotected base pairs;
- (d) providing a preselected site from which to sequence;
- and sequencing the sample polynucleotide strands to identify the alteration(s).
2 more elements
Dependencies are inferred from the claim text.