Methods for obtaining nucleic acid containing a mutation
First Claim
1. A method for preparing a heteroduplex-enriched nucleic acid sample from an organism carrying heterozyygous mutations, comprising the steps of:
- (a) denaturing substantially all of the double-stranded nucleic acid present in a nucleic acid sample from an organism;
(b) allowing the nucleic acid to anneal the under conditions which permit formation of heteroduplexes and homoduplexes; and
(c) removing homoduplexes from the annealed sample wherein the removal of said homoduplexes is effected using immobilized mismatch binding proteins (MBP), thereby retaining heteroduplexes in said sample.
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Abstract
In order to facilitate the screening of an organism, or a population of organisms, carrying heterozygous mutations for identifying the presence of a mutation in a gene of interest, a method is provided which utilizes mismatch binding proteins, such as MutS. The method comprises the steps of denaturing double stranded nucleic acid present in a nucleic acid sample from an organism; allowing the nucleic acid to anneal; and removing homoduplexes from the annealed sample, thereby retaining heteroduplexes in the sample. A positive signal from a probe specific for the gene of interest indicates that the organism carries a mutation in the gene.
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Citations
10 Claims
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1. A method for preparing a heteroduplex-enriched nucleic acid sample from an organism carrying heterozyygous mutations, comprising the steps of:
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(a) denaturing substantially all of the double-stranded nucleic acid present in a nucleic acid sample from an organism; (b) allowing the nucleic acid to anneal the under conditions which permit formation of heteroduplexes and homoduplexes; and (c) removing homoduplexes from the annealed sample wherein the removal of said homoduplexes is effected using immobilized mismatch binding proteins (MBP), thereby retaining heteroduplexes in said sample. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
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Specification