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196 results about "Genetic typing" patented technology

Construction method and sequencing method of genetic typing sequencing library

The invention discloses a construction method and a sequencing method of a genetic typing sequencing library. The construction method comprises the following steps: S1, bonding sample label sequences at the two ends of a first enzyme digestion fragment while a gene group DNA is subjected to enzyme digestion for the first time to produce the first enzyme digestion fragment so as to obtain a first enzyme digestion fragment with a label; S2, performing PCR amplification on the first enzyme digestion fragment by utilizing an amplification primer with a library label sequence to obtain the gene typing sequencing library. According to the construction method, the sample label sequences are directly bonded at the two ends of the first enzyme digestion fragment while the gene group DNA is subjected to enzyme digestion for the first time to produce the first enzyme digestion fragment, so that the constructed library can directly get into the next step of reading a target enzyme digestion fragment sequence after the sample label information is read during sequencing by a computer, and the reading length of the target enzyme digestion fragment sequence is increased, the invalid data volume is reduced, and the effective quantity of sequencing data is increased.
Owner:BEIJING NOVOGENE TECH CO LTD

Molecular markers closely linked with multiple-effect major quantitative trait loci (QTL) of grain weight or silique length of rape, and application thereof

The invention discloses molecular markers closely linked with multiple-effect major QTL of the grain weight or silique length of rape, and application thereof. The closely-linked molecular markers canbe used for molecular marker-assisted selection of rape and map-based cloning of the QTL. According to the invention, a hybrid F1 and parents are subjected to continuous backcrosses for multiple generations, and a hybrid near-isogenic line is constructed by cooperatively using a molecular marker-assisted selection method; individual plants with high background reversion rate are subjected to selfing so as to obtain a QTL-NIL segregation population; molecular markers are gradually encrypted in a target region so as to realize genotype analysis of the population and searching of crossover individual plants; and thus, the molecular markers BrSF7-101 and BrSF6-2389 are obtained, and the physical distance between the two molecular markers is only 134 kb. The two closely-linked molecular markers are used for genetic typing of linked populations, and it is found that two alleles are greatly different in grain weight and silique length; and thus, the molecular markers can be applied to molecular marker-assisted selection of rape, improve selection efficiency and accelerate breeding progress.
Owner:INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI

KIR and ligand genetic typing experimental method

The invention discloses a KIR and ligand genetic typing experimental method. By the technology, the time and labor are saved, and meanwhile, DNA sample capacity is further saved. A multi-PCR technology is adopted, meanwhile, 36 pairs of primers are further combined according to the sizes of PCR products, and finally, amplified reaction is finished in 12 reaction holes. The KIR and ligand genetic typing experimental method is conveniently applied to amplification of 96 pore plates, conventional Taq enzyme is used, typing of all KIR genes and ligands thereof can be finished at a time under the same reaction conditions, and the practicality of the KIR and ligand genetic typing experimental method is greatly improved. Two pairs of primers are used for a KIR gene, the accuracy is improved, anda false negative result is avoided. The KIR gene can be combined to MHC-I type ligand molecules on the surfaces of targeting cells, inhibiting or activating signals are transmitted to regulate the activity of NK cells and T cells, and the KIR and ligand genetic typing experimental method plays an important regulation role in hematopoietic stem cell transplantation, feto-matemal tolerance, anti-infectious immunity, tumor immunity and autoimmune diseases. Therefore, KIR genetic typing facilitates understanding of influences of KIR to tumor immunity, hematopoietic stem cell transplantation and autoimmune diseases.
Owner:韩瑜

Mitochondrial SNP fluorescence labeling composite amplification kit and application thereof

The invention relates to a mitochondrial SNP fluorescence labeling composite amplification kit capable of simultaneously detecting 35 SNP sites and main application thereof. The kit employs 35 pairs of specific primers for genetic typing of human mitochondrial SNP sites. The kit comprises a PCR amplification reaction reagent, FAM color composite amplification primers, HEX color composite amplification primers, a positive control, a negative control and fluorescent internal standard. An application method of the kit comprises the following steps: carrying out PCR composite amplification on mitochondrial DNAs extracted from a variety of to-be-detected materials; and collecting fluorescence signals through electrophoresis by using a genetic analyzer, which enables accurate allele typing of each site to be realized. Compared with a conventional mitochondrial SNP sequencing method, the kit provided by the invention is more simple, convenient and economic, uses less templates and has a higher success rate. As a mitochondrial detection system, the kit carries out simultaneous amplification in two tubes and simultaneous electrophoresis, diversity of haplotype reaches 99.64%, and the kit can be used as a kit for identification of a same maternal line following an autosomal STR and a Y chromosome STR.
Owner:SHANG HAI GENE BIO TECH +2

Time-of-flight mass spectrometry nucleic acid analysis method for detecting human spinal muscular atrophy gene mutation

The invention discloses a time-of-flight mass spectrometry nucleic acid analysis method for detecting human spinal muscular atrophy gene mutation. A primer combination comprising an amplification primer and a mass spectrometry extension probe primer is utilized. According to the method, the copy numbers of sequences related to SMN1, SMN2, NAIP, H4F5 and GTF2H2 genes are quantitatively detected, and whether deletion, deletion number and multiple copies exist or not is analyzed, so that the clinical phenotype severity can be directly deduced; the method has good sensitivity, specificity, stability and accuracy, and effectively solves the technical bottleneck of false negative, false positive and the like; the operation is simple, the cost is relatively low, and the result is stable and reliable; the method is high in flux and low in cost, has general representativeness and universality, is easy to realize automatic and large-scale detection, and is suitable for large-scale population screening; genetic typing detection can be carried out on part of common SMN1 upper point mutations; and the requirements of large-scale population screening, prenatal diagnosis and conventional molecular diagnosis in current SMA prevention and treatment are met.
Owner:GUANGZHOU DARUI BIOTECH

Multi-gene pyramiding breeding method for thoroughbred milk goats

The invention discloses a multi-gene pyramiding breeding method for thoroughbred milk goats. Genome DNA (deoxyribonucleic acid) of a milk goat continuously giving birth to two or more lambs is used as a template, four pairs of primers respectively expand intron 2 and exon 10 of a prolactin receptor gene and an untranslation region (5'UTR) and exon 1 of luteotropin beta calcmeurin 5', the sizes of expanded products are judged by agarose gel electrophoresis, site mutation of the expanded products of the four pairs of primers are screened by DNA sequencing technology, then polyacrylamide gel electrophoresis is used for performing genetic typing and gene frequency analysis for SNPs (single nucleotide polymorphisms) of four sites of the prolactin receptor gene and the luteotropin beta calcmeurin, the relation of polymorphism of a multiple-birth milk goat individual (F1 generation) and the number of born lamps and the relation of different genetype combinations and the number of the born lamps are analyzed, parental generation (F0 generation) is reviewed, filial generation (F2 generation) is tracked, the relation of the genetype combination of an ewe individual and the number of the born lamps is detected, and contribution of different genetypes in terms of prolific trait formation is analyzed.
Owner:NORTHWEST A & F UNIV

Method for simultaneously measuring gene polymorphism of tacrolimus action targets of person

The invention belongs to the technical field of molecular biology and medical examination, and relates to a matrix-assisted laser desorption ionization flight time mass spectrum method for simultaneously measuring signal channel series protein coding gene polymorphism of tacrolimus action targets of a person. The method comprises the following steps of performing multiple PCR (polymerase chain reaction) amplification after performing DNA (deoxyribonucleic acid) extraction on a sample; and performing genetic typing analysis on the sample by using a flight time mass spectrum method after single nucleotide primer extension. The method is easy and convenient to operate, a sequence specificity probe is not required, multiple detection on a plurality of samples can be implemented, 40 reactions can be realized by each system, 40 SNP (single nucleotide polymorphism) sites can be detected simultaneously, throughput is high, application range is wide, and tens of thousands of samples and dozens of to hundreds of sites can be detected simultaneously; fluorescent markers are not required, only common primers require to be synthesized, and analysis cost of a single sample is low; and flexibility is high, the amount of samples required for analysis is small, and sensitivity and detection precision are high. The method is suitable for detecting clinic conventional genes, and further provides a technical support for clinic individual rational drug use.
Owner:AFFILIATED HUSN HOSPITAL OF FUDAN UNIV
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