58 results about "Allele-specific oligonucleotide" patented technology

The present invention provides for methods for discriminating between alleles at polymorphic positions in a genome. In general the methods employ allele specific extension of oligonucleotides that are complementary to one of the alleles at the 3′ end of the oligonucleotide. The allele specific oligonucleotides are resistant to proof reading activity from a polymerase and may be extended in an allele specific manner by a DNA polymerase with a functional 3′ to 5′ exonuclease activity. The allele specific oligonucleotides may be attached to a solid support such as a chip or a bead.

The present invention includes a method of allele-specific amplification, utilizing an allele-specific oligonucleotide, at least partially complementary to more than one variant of the target sequence, but having a 3′-terminal nucleotide complementary to only one variant of the target sequence and having at least one nucleotide with a base covalently modified at the exocyclic amino group, wherein the allele-specific oligonucleotide is extended by a nucleotide-incorporating biocatalyst predominantly when hybridized to the variant of the target sequence for which it has said complementary 3′-terminal nucleotide.

The invention encompasses methods and products related to genotyping. The method of genotyping of the invention is based on the use of single nucleotide polymorphisms (SNPs) to perform high throughput genome scans. The high throughput method can be performed by hybridizing SNP allele-specific oligonucleotides and a reduced complexity genome (RCG). The invention also relates to methods of preparing the SNP specific oligonucleotides and RCGs, methods of fingerprinting, determining allele frequency for a SNP, characterizing tumors, generating a genomic classification code for a genome, identifying previously unknown SNPs, and related compositions and kits.

The present invention discloses the use of Allele-Specific-oligonucleotide (ASO) as a detection assay for human HLA classification. Using Reversed-Dot-Blotting format and flow through hybridization process, more efficient, faster and less expensive HLA classification can be achieved. A simplified procedure for HLA genotyping is also described. This invention further provides a Single Nucleotide Polymorphism (SNP)-based DNA fingerprining method for rapid and accurate genotyping, identification as well as DNA analyses of genetic data from human beings and different organisms. In addition this invention also discloses a new device for rapid and sensitive analyses of nucleic acids, proteins and other analysts for diagnosis.

The present invention disclosed the use of single nucleotide polymorphism (SNP) as the detection assay for human identification. Using the reversed dot-blot format and the flow through hybridization process, the process can be more efficient, less expensive and with similar or better power of exclusion in definitive identification. The present method can be applied to any other organisms.

A method of genotyping blood cell antigens comprising subjecting DNA from an individual of a mammalian species to a multiplex Polymerase Chain Reaction (PCR) to amplify and detectably label a region of the locus of at least two different blood cell antigens which contains the site of nucleotide polymorphism of said blood cell antigen and using the thus amplified and labeled DNA fragments to determine the genotype for each of said blood cell antigens. The multiplex PCR comprises the use of at least one pair of blood cell antigen-specific chimeric primers for each blood cell antigen to be genotyped and at least one detectably labeled universal primer, preferably a pair of detectably labeled universal primers. The universal primer(s) have a unique sequence not occurring in the DNA of said mammalian species. Each chimeric primer pair comprises a left chimeric primer and a right chimeric primer, each of them comprising a blood cell antigen-specific part at the 3' end and a universal part at the 5' end. The base sequence of the universal part of the chimeric primers corresponds to the base sequence of said at lea st one universal primer. The blood cell antigen-specific parts of the chimeric primer pair enclose a region of the locus of the blood cell antigen which contains the site of nucleotide polymorphism of said blood cell antigen. A k it for genotyping blood cell antigens by this method. A set of blood cell antig en- specific chimeric primer pairs and a set of blood cell antigen allele-specif ic oligonucleotide probes.

A method of genotyping blood cell antigens comprising subjecting DNA from an individual of a mammalian species to a multiplex Polymerase Chain Reaction (PCR) to amplify and detectably label a region of the locus of at least two different blood cell antigens which contains the site of nucleotide polymorphism of said blood cell antigen and using the thus amplified and labeled DNA fragments to determine the genotype for each of said blood cell antigens. The multiplex PCR -comprises the use of at least one pair of blood cell antigen-specific chimeric primers for each blood cell antigen to be genotyped and at least one detectably labeled universal primer, preferably a pair of detectably labeled universal primers. The universal primer(s) have a unique sequence not occurring in the DNA of said mammalian species. Each chimeric primer pair comprises a left chimeric primer and a right chimeric primer, Each of them comprising a blood cell antigen-specific part at the 3′ end and a universal part at the 5′ end. The base sequence of the universal part of the chimeric primers corresponds to the base sequence of said at least one universal primer. The blood cell antigen-specific parts of the chimeric primer pair enclose a region of the locus of the blood cell antigen which contains the site of nucleotide polymorphism of said blood cell antigen. A kit for genotyping blood cell antigens by this method. A set of blood cell antigen-specific chimeric primer pairs and a set of blood cell antigen allele-specific oligonucleotide probes.

The present invention includes a method of allele-specific amplification, utilizing an allele-specific oligonucleotide, at least partially complementary to more than one variant of the target sequence, having an internally-placed selective nucleotide complementary to only one variant of the target sequence wherein the allele-specific oligonucleotide is extended by a nucleic acid polymerase predominantly or exclusively when hybridized to the variant of the target sequence for which it has said complementary selective nucleotide.

The present invention includes a method of allele-specific amplification, utilizing an allele-specific oligonucleotide, at least partially complementary to more than one variant of the target sequence, but having a 3′-terminal nucleotide complementary to only one variant of the target sequence and having at least one nucleotide with a base covalently modified at the exocyclic amino group, wherein the allele-specific oligonucleotide is extended by a nucleotide-incorporating biocatalyst predominantly when hybridized to the variant of the target sequence for which it has said complementary 3′-terminal nucleotide.

The present invention includes a method of allele-specific amplification, utilizing an allele-specific oligonucleotide, at least partially complementary to more than one variant of the target sequence, but having at least one selective nucleotide complementary to only one variant of the target sequence and incorporating at least one “G-clamp” nucleotide.

The invention discloses a flight mass spectrometry genotyping detection method. The method comprises the following steps: (1) designing three oligonucleotide probes for a SNP site, wherein the 3 / endsof a first probe and a second probe are respectively two allelic genes, i.e., allele-specific oligos (ASOs) of a pre-selected SNP site, (2) when the sequence is completely matched with the target sequence, preparing a connection product, (4) carrying out enzyme digestion on the PCR product by using restriction endonuclease, and purifying the product by using streptavidin-coated beads, and (5) transferring the purified product to a 384-well chip. SNP site typing is carried out by adopting MALDI-TOF mass spectrometry, and SNP typing is provided by adopting the method which is simple to operate,low in cost and high in resolution ratio.

The invention belongs to the field of biology, and relates to single base mutation. More particularly, the invention relates to a mutation site of a 293th site of SEQ ID NO.2 corresponding to a CYP2C9 gene, wherein the mutation site is mutated into T from wild type G. The invention also relates to a nucleic acid segment, a coded protein segment and application thereof. The invention also provides an allele-specific oligonucleotide, kit and method for detecting the mutation site.

The invention belongs to the field of biology, relates to single-base mutation and particularly relates to a 1300th bit mutation site of a CYP2C9 gene corresponding to SEQ ID NO.2. The 1300th bit mutation site is mutated from wild type A into T. The invention relates to a nucleic acid fragment containing the 1300th bit mutation site, a protein fragment encoded through the same and application thereof. The invention also provides allelomorphic gene specific oligonucleotide for detecting the 1300th bit mutation site, a kit and a detection method.

The invention belongs to the field of biology, and relates to single base mutation. More particularly, the invention relates to a mutation site of a 679th site of SEQ ID NO.2 corresponding to a CYP2C9 gene, wherein the mutation site is mutated into T from wild type C. The invention also relates to a nucleic acid segment, a coded protein segment and application thereof. The invention also provides an allele-specific oligonucleotide, kit and detection method for detecting the mutation site.

The invention belongs to the field of biology, and relates to single base mutation. More particularly, the invention relates to a mutation site of a 287th site of SEQ ID NO.2 corresponding to a CYP2C9 gene, wherein the mutation site is mutated into C from wild type G. The invention also relates to a nucleic acid segment, a coded protein segment and application thereof. The invention also provides an allele-specific oligonucleotide, kit and detection method for detecting the mutation site.

The invention belongs to the field of biology, relates to single-base mutation, and in particular relates to a CYP2C9 gene which is corresponding to the mutation site mutated from wild type C into A on the 1009th bit of SEQ ID NO.2, a nucleic acid segment containing the mutation site, a protein segment coded by the nucleic acid segment and application thereof. The invention also provides allele specific oligonucleotide, a kit and a detection method for detecting the mutation site.

The invention belongs to the field of biology, and relates to single base mutation. More particularly, the invention relates to a mutation site of a 448th site of SEQ ID NO.2 corresponding to a CYP2C9 gene, wherein the mutation site is mutated into T from wild type C. The invention also relates to a nucleic acid segment, a coded protein segment and application thereof. The invention also provides an allele-specific oligonucleotide, kit and detection method for detecting the mutation site.

The invention belongs to the field of biology, and relates to single base mutation, and more specifically, the invention relates to a mutation site, corresponding to the 637th site of SEQ ID NO.2, ofCYP2C9 gene, the site is mutated from wild type A into G, a nucleic acid fragment containing the mutation site, a protein fragment coded by the nucleic acid fragment and application of the nucleic acid fragment are disclsoed. The invention also provides allele-specific oligonucleotides, kits and detection methods for detecting the mutation site.

The invention belongs to the field of biology, and relates to single base mutation, and more specifically, the invention relates to a mutation site, corresponding to the 419th site of SEQ ID NO.2, ofCYP2C9 gene; the site is mutated into A from wild-type G, a nucleic acid fragment containing the mutation site, a protein fragment coded by the nucleic acid fragment and application of the nucleic acid fragment are disclosed. The invention also provides allele-specific oligonucleotides, kits and detection methods for detecting the mutation site.

The invention belongs to the field of biology, relates to single-base mutation, and in particular relates to a CYP2C9 gene which is corresponding to the mutation site mutated from wild type T into C on the 1400th bit of SEQ ID NO.2, a nucleic acid segment containing the mutation site, a protein segment coded by the nucleic acid segment and application thereof. The invention also provides allele specific oligonucleotide, a kit and a detection method for detecting the mutation site.

The present invention includes a method of allele-specific amplification, utilizing an allele-specific oligonucleotide, at least partially complementary to more than one variant of the target sequence, but having at least one selective nucleotide complementary to only one variant of the target sequence and incorporating both a nucleotide with a base covalently modified at the exocyclic amino group and a modified phosphate.

The invention belongs to the field of biology, and relates to single base mutation. More particularly, the invention relates to a mutation site of a 394th site of SEQ ID NO.2 corresponding to a CYP2C9 gene, wherein the mutation site is mutated into T from wild type C. The invention also relates to a nucleic acid segment, a coded protein segment and application thereof. The invention also provides an allele-specific oligonucleotide, kit and method for detecting the mutation site.

A method for genotyping a subject with respect to a gene or target nucleic acid sequence associated with a pathological condition, said method comprising contacting an allele specific oligonucleotide immobilized to a solid support with a single-stranded from of RNA or DNA from a subject to be tested labeled directly or indirectly with a reporter molecule capable of giving an identifiable signal under conditions which permit hybridization of single stranded RNA or DNA which is exactly complementary to the immobilized allele specific oligonucleotide but substantially less or no hybridization of non-complementary single-stranded RNA or DNA molecules and then screening for the presence or absence or level of reporter molecule which provides an indicator of the genetic identity of the single-stranded RNA or DNA molecule which in turn provides the genotype of the subject.

The present invention is generally drawn to a novel method for determining a SNP (single nucleotide polymorphism) genotype using a PTO-SNV (Probing and Tagging Oligonucleotide for Single Nucleotide Variation). The present invention provides novel protocols for SNP genotyping in which only one allele-specific oligonucleotide permits in a SNP genotyping reaction to determine whether a target nucleic acid sequence to be analyzed is homozygous or heterozygous for the SNP allele of interest or has no SNP allele of interest.