151 results about "High Resolution Melt" patented technology

The invention provides a preparation method of a non-transgenic mutant based on a CRISPR-Cas9 technology. Specifically, the preparation method comprises a construction method and a screening method. The construction method is characterized in that CRISPR-Cas9 and sgRNA are preassembled and are located at T-DNA of agrobacterium tumefaciens; site-directed change of a target gene of a target plant can be realized by infection of the target plant by the agrobacterium tumefaciens and coculture without integrating the sequences of the CRISPR-Cas9 and the sgRNA into a genome of the target plant, so that an obtained site-directed mutant material of the target gene does not need the processes of sexual reproduction, segregation posterity, population screening and the like or does not contain any exogenous gene sequence. The obtained regeneration seedlings are screened by a high-throughput sequencing and high-resolution melting curve technology provided by the invention; mutant plants of which target genes are mutated can be obtained by identifying the regeneration seedlings efficiently and quickly at the current generation of transgene, even if low-proportion mutants of which the proportionis 1 / 100 of a mixed population can be screened. The screening method provided by the invention still can ensure excellent accuracy and sensitivity.

The invention relates to the gene detecting field, especially a method for detecting gene copy number variation. The method comprises: selecting an endogenous or exogenous similar sequence which is similar to and different from the gene sequence to be detected; designing a corresponding a primer for amplifying the gene sequence to be detected and the similar sequence simultaneously, then performing the high-resolution melting curve analysis; when there is copy number variation in the gene to be detected, the result of the high-resolution melting curve analysis can be clearly distinguished from the result of the analysis of the wild-type gene to be tested, so as to realize the detecting target. The method of the invention is not only suitable for gene copy number variation, which is equally suitable for aneuploidy detection. The method of the invention is not only suitable for gene copy number variation, population screening and conventional prenatal diagnosis of aneuploidy detection, which is further equally suitable for non-invasive prenatal diagnosis. The method of the invention has advantages of rapidness, precision, high sensitivity, high throughput, low cost and no pollution.

The invention relates to a method and a kit for detecting gene mutation, in particular to a method and a kit for detecting the mutation of BRAF gene. The invention is characterized in that the kit comprises a specific probe used for carrying out genotyping on No. 15 exon codon V600E of the BRAF gene, wherein the specific probe of the No. 15 exon codon V600E comprises a nucleotide sequence of V600E codon. by the technology combining the conventional polymerase chain reaction (PCR) amplification with a Cold-PCR enrichment amplification product and a high resolution melting curve analysis technology, the kit provided by the invention can be used for completing the judgment on sample genotyping.

The invention discloses a method for detecting relative gene polymorphism of warfarin personalized medication by using a high-resolution melting curve analysis technology. The method is characterized by comprising the following steps: adopting a silica gel adsorption method to extract genome DNA (Deoxyribose Nucleic Acid) of an oral epithelial cell / peripheral blood cell sample of a detected person; designing a detection primer containing a target site; carrying out site typing on a PCR (Polymerase Chain Reaction) amplified gene sequence by using the high-resolution melting curve (HRM) analysis technology. The method disclosed by the invention is high in sensitivity, good in specificity and fast in detection speed, and can be used for providing reference to a clinical medication dosage of warfarin, so as to realize reasonable and effective personalized medication.

The present invention relates to a Polymerase Chain Reaction and High Resolution Melt genetic identification system, and, more specifically, to a tactical and portable Polymerase Chain Reaction and High Resolution Melt genetic analysis and identification system that is configured to determine and communicate analysis and identification results and a tiered confidence / alert level related to the analysis and identification results.

The invention provides a method for detecting an FOXH1 gene SNP locus rs750472 genotype. The method comprises the following steps: (1) constructing a recombinant negative plasmid and a recombinant positive plasmid; (2) extracting a peripheral blood genomic DNA of a to-be-detected sample, carrying out PCR amplification and HRM analysis of the recombinant negative plasmid, the recombinant positive plasmid and the peripheral blood genomic DNA of the to-be-detected sample, and collecting data; and (3) drawing high resolution melting curves according to the collected data, and judging the FOXH1 gene SNP locus rs750472 genotype of the to-be-detected sample according to the melting curves of the recombinant negative plasmid and the recombinant positive plasmid. The method can effectively detect the FOXH1 gene SNP locus rs750472 genotype, and has the advantages of high sensitivity, good specificity, and rapid detection; and an analysis result with application of the method is entirely consistent to a sequencing result, and the method can be used for assistant judgment of ventricular septal defect heart disease.