Mutation Analysis
AssigneeEpistem Limited
InventorBen COBB
Life of this patent
Estimating the term…
Abstract
A method and an oligonucleotide probe are described for determining the presence or absence of mutant alleles in a genomic locus. The probe binds to different alleles of a target sequence with different melting temperatures (Tm). The method determines the Tm of the probe when it is hybridized to the target sequence to establish whether a variant nucleic acid such as a mutant allele is present or absent in the target sequence. There may be variants in a target sequence that are not of interest, for example phenotypically silent mutations. To ensure that these variants do not influence the Tm of the probe, the probe contains universal base sites where such variants of no interest occur.
Claims27
A method for detecting the presence of a variant nucleic acid sequence in a target nucleic acid sequence that is polymorphic in that it is present in multiple alleles within a given population, the method comprising
- a) providing a reaction mix comprising:
- an oligonucleotide probe having a first melting temperature (Tm) when hybridised to a first allele of the target sequence, and a second lower Tm when hybridised to a second allele of the target sequence, wherein the probe comprises at least one universal base at a site where variants are not desired to be detected;
- and
- a sample comprising nucleic acid whose nucleotide sequence comprises the target sequence;
- b) allowing the probe to hybridise to nucleic acids in the sample;
- and
- c) determining the Tm of the probe when hybridised to the nucleic acids in the sample;
- to thereby determine whether a variant nucleic acid sequence is present in the sample.
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An oligonucleotide probe having a first melting temperature (Tm) when hybridised to a first allele of a target sequence, and a second lower Tm when hybridised to a second allele of a target sequence, wherein the probe comprises at least one universal base at a site where variants are not desired to be detected.
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Dependencies are inferred from the claim text.