35 results about "Rare mutations" patented technology

Many areas of biomedical research depend on the analysis of uncommon variations in individual genes or transcripts. Here we describe a method that can quantify such variation at a scale and ease heretofore unattainable. Each DNA molecule in a collection of such molecules is converted into a single particle to which thousands of copies of DNA identical in sequence to the original are bound. This population of beads then corresponds to a one-to-one representation of the starting DNA molecules. Variation within the original population of DNA molecules can then be simply assessed by counting fluorescently-labeled particles via flow cytometry. Millions of individual DNA molecules can be assessed in this fashion with standard laboratory equipment. Moreover, specific variants can be isolated by flow sorting and employed for further experimentation. This approach can be used for the identification and quantification of rare mutations as well as to study variations in gene sequences or transcripts in specific populations or tissues.

The present invention is directed to a method for detecting and quantifying rare mutations in a nucleic acid sample. The nucleic acid molecules under investigation can be either DNA or RNA. The rare mutation can be any type of functional or non-functional nucleic acid change or mutation, such as deletion, insertion, translocation, inversion, one or more base substitution or polymorphism. Therefore, the methods of the present invention are useful in detection of rare mutations in, for example, diagnostic, prognostic and follow-up applications, when the targets are rare known nucleic acid variants mixed in with the wildtype or the more common nucleic acid variant(s).

The present invention is directed to a method for detecting and quantifying rare mutations in a nucleic acid sample. The nucleic acid molecules under investigation can be either DNA or RNA. The rare mutation can be any type of functional or non-functional nucleic acid change or mutation, such as deletion, insertion, translocation, inversion, one or more base substitution or polymorphism. Therefore, the methods of the present invention are useful in detection of rare mutations in, for example, diagnostic, prognostic and follow-up applications, when the targets are rare known nucleic acid variants mixed in with the wildtype or the more common nucleic acid variant(s).

The invention relates to a microcavity multicolor fluorescent digital PCR chip and application thereof. The chip comprises a liquid inlet port, a double side microcavity array and a double side wasteliquid tank. The chip is used for rare cancer rare mutation analysis, differential gene expression analysis or high precision copy number variation analysis. The multicolor fluorescence detection is used, so that the detection flux of a single chip can be greatly improved, the reaction efficiency is greatly improved, the operation is convenient, the speed is high, extremely high detection sensitivity can be achieved compared with an existing real-time fluorescence quantitative PCR technology, and the application prospect is good.

Loop-shaped primer used in nucleic acid amplification is an oligonucleotide with 3-20 bases in both 3′ and 5′ ends which can be combined together to form a double-strand under appropriate conditions, resulting in the primer forming a stem-loop structure. The double-stranded structure is opened and the stem-loop structure dissolves when the primer recognizes and hybridizes with the target sequence. If the target sequence is not present the primer can form a stem-loop structure automatically by self-annealing. The primer can comprise a universal tag sequence or not. Together with universal tag sequence primer the primer comprising the universal tag sequence can be used for a second round of amplification. The primer has high specificity and does not form a primer dimmer. The primer is easy to design and is suitable for measuring gene expression and detecting features of nucleic acids such as SNPs and rare mutations.

The invention discloses a super-blocking fluorescent quantitative PCR method for detecting rare mutations with high sensitivity. The invention adopts the technology of specific primers and probes, and can realize rapid detection of gene point mutations in blood samples. The advantages of the present invention include: (1) On the basis of conventional ARMS primers, LNA modification is added or additional mismatched bases are introduced at the last two to three positions of the 3', which does not affect the sensitivity of mutation detection and ensures the specificity of mutations (2) Introduce a modified blocking probe that is completely complementary to the wild type, and further reduce the background interference of the wild type on the basis of taking into account the amplification efficiency of the mutant type; (3) After introducing a highly specific On the basis of non-modified blocking probes, ARMS primers can detect different base mutations at the same site at the same time, so as to realize the merger detection of multiple sites in the same gene with only a few systems; (4) high sensitivity; (5) The detection speed is fast.

The invention relates to a genetic detection method, in particular to a quantitative detection method for unusual genetic mutation. The quantitative detection method for unusual genetic mutation comprises the following steps: firstly, corresponding primer sequences are designed and prepared according to the unusual genetic mutation required for quantitative detection, wherein the 3' end of a primer is provided with a mutant site - special nucleotide; except for the final site of the nucleotide, all the other sequences can be synthesized by commercial means; and the final site of the nucleotide is modified nucleotide, so as to make the primer incapable of extending in the general PCR reaction; and the modified nucleotide is added to the tail of the primer; and secondly, in a reaction system which contains special DNA polymerase and pyrophosphate, the primer designed and prepared in the first step is adopted for amplifying the unusual genetic mutation for quantitative detection, and a real-time fluorescent PCR instrument is utilized for quantitative detection; and the quantitative detection information of the unusual genetic mutation can be acquired through analysis of an amplifying curve recorded by the real-time fluorescent PCR instrument.

The present disclosure encompasses methods of error corrected sequencing (ECS) that enable detection of very rare mutations well below the error rate of convention next generation sequencing (NGS). Further, the methods disclosed herein enable multiplex targeting of genomic DNA.