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US 5,633,134 AGranted
Method for simultaneously detecting multiple mutations in a DNA sample
AssigneeIG Laboratories, Inc.
InventorTony Shuber
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Abstract
The invention provides a rapid, cost-effective process for simultaneously testing large numbers of individuals for the presence or absence of multiple mutaitons in one gene or multiple genes using allele specific oligonucleotide (ASO) probes in the presence of a quaternary ammonium salt which eliminates disparities in the melting temperatures of the ASO used.
Claims20
A method of screening to simultaneously detect multiple target sequences of interest in genomic DNA, the method comprising:
- (a) immobilizing on a single solid support a sample of genomic DNA to be screened for said multiple target sequences of interest,
- (b) contacting said support with a pool of oligonucleotide probes of approximately the same length under stringent conditions in the presence of a concentration of about 2-5 M of a quaternary ammonium salt that allows the hybridization of said probes to DNA under said stringent conditions,
- wherein said pool comprises individual allele-specific oligonucleotide (ASO) probes, each ASO complementary to and specific for one of said multiple target sequences of interest suspected to be present in said DNA;
- (c) removing probes that do not hybridize to said DNA under said stringent conditions;
- and
- (d) detecting hybridization, wherein said hybridization indicates the presence of at least one of said multiple target sequences of interest in said DNA.
3 more elements
A method of screening to simultaneously detect multiple target sequences of interest in genomic DNA, the method comprising:
- (a) immobilizing on a single solid support a sample of genomic DNA to be screened for said multiple target sequences of interest,
- (b) contacting said support with a pool of oligonucleotide probes of approximately the same length under stringent conditions in the presence of a concentration of about 2-5 M of a quaternary ammonium salt that allows the hybridization of said probes to DNA under said stringent conditions,
- wherein said pool comprises individual allele-specific oligonucleotide (ASO) probes, each ASO complementary to and specific for one of said multiple target sequences of interest suspected to be present in said DNA;
- (c) removing probes that do not hybridize to said DNA under said stringent conditions,
- (d) detecting hybridization, wherein said hybridization indicates the presence of at least one of said multiple target sequences of interest in said DNA,
- (e) contacting said DNA having at least one of said multiple target sequences of interest with an individual ASO probe under stringent conditions, wherein said contacting occurs in the presence of a concentration of a quaternary ammonium salt that allows the hybridization of said probe to DNA under said stringent conditions, and
- (f) detecting hybridization, wherein said hybridization indicates the presence of a target sequence complementary to said individual ASO probe in said DNA.
4 more elements
Dependencies are inferred from the claim text.