196 results about "Genetic typing" patented technology

The invention discloses a fluorescence detection kit capable of detecting 17 non-syndromic hereditary hearing loss susceptibility genes. The kit adopts 17 pairs of specific primers to conduct genetic typing on the hearing loss susceptibility genes, and 17 hotspot mutations in four most common Chinese hearing loss related genes can be detected at the same time in a single tube within 3 hours. The kit comprises primer combinations of 17 polymorphic sites of hereditary hearing loss on a GJB2 (CX26) gene, an SLC26A4 (PDS) gene, a GJB3 gene and a 12SrRNA (MTRNR1) gene, can be used for accurately judging the wild type, the pure mutant type or the hybrid type of the 17 sites, and achieves diagnosis and screening of the hearing loss genes. The kit provided by the invention can be applied to rapidly and efficiently detecting the hearing loss genes and is a rapid, convenient, economical and efficient screening kit for hearing loss virulence genes.

The invention belongs to the technical field of forensic medicine, and particularly relates to a forensic medicine II sequence testing kit based on 74 gama chromosome SNP genetic markers. The technical problem to be solved is to classify detection materials of Chinese group source at the position of a gama chromosome evolution tree by using the gama chromosome SNP genetic markers. The technical scheme of the invention is forensic medicine II sequence testing kit based on 74 gama chromosome SNP genetic markers, which includes a mixture of 72 pairs of primers for recombination and amplification, thus the detection of 74 pieces of SNP at the same time becomes true. The kit applies single pipe internal recombination and amplification and II sequence testing technology, and thus the genetic typing of 74 pieces of gama chromosome SNP genetic markers of multiple biological detection materials can be obtained in one time; besides, male samples of Chinese group source are correctly affiliated to the branch of the recognized gama chromosome evolution tree.

The invention discloses a fast and efficient saliva DNA extraction kit and an extraction method. The fast and efficient saliva DNA extraction kit contains cell lysis buffer, DNA combined liquid, rinsing liquid and eluant. The extraction method of saliva DNA at least comprises the following three steps: (1) cell lysis: adding the cell lysis buffer into saliva, thereby obtaining the cell lysis buffer containing free DNA; (2) adding DNA combined liquid into the cell lysis buffer obtained in the step (1), thereby obtaining a DNA-magnetic bead absorbing compound; (3) utilizing rinsing liquid and eluant to elute the compound generated in the step (2), and removing RNA and other impurities, thereby obtaining high-purity DNA. The novel cell lysis buffer is adopted by the fast and efficient saliva DNA extraction kit; the cost is low; the extracted DNA purity is high; the completeness is excellent; no RNA is remained; the fast and efficient saliva DNA extraction kit can be applied to high-throughput sequencing, genetic typing and general molecular biological experiment operation.

The invention provides a method, oligonucleotide and kit for detecting common high-risk HPV (human papilloma viruses). A fluorescence PCR (polymerase chain reaction) technology is adopted, HPV16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 68, 26, 53, 66, 73 and 82 which possibly exist in a sample are subjected to initial detection and type identification. By the method, oligonucleotide and kit for detecting the common high-risk HPV, 18 high-risk types can be detected simultaneously, and the HPV 16 and 18 can be subjected to genetic typing simultaneously. On the basis of 18 types of high-risk HPV and affinity of other types on a phylogenetic tree, a high-risk HPV primer and a probe are designed, a background fluorescence value is reduced remarkably while detection accuracy, sensitivity and specificity are guaranteed, detection flux is increased remarkably, and detection cost is greatly reduced.

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.

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.

The invention provides a sequencing primer for qualitative detection of cytochrome oxidase CYP2C19 genetic typing and a kit of the sequencing primer and belongs to the field of external nucleic acid detection. The kit comprises uracil DNA (Deoxyribonucleic Acid) glycosylase, Taq polymerase, PCR (Polymerase Chain Reaction) reaction liquor, PCR amplification primers, pyrophosphoric acid sequencing primers and positive control products. The kit is high in sensitivity and good in specificity, PCR products can be simply treated for a pyrophosphoric acid sequenator for sequencing, operation is simple, reaction time is short, the sensitivity of the kit is higher than that of a golden standard-capillary electrophoresis sequencing, and the kit is suitable for use for mutation analysis.

The invention belongs to the field of molecular genetics and particularly relates to a molecular marking method of a mutant site related to a 5' control region of a chicken miR-1b-3p gene and chicken sexual maturity and laying number traits and an application thereof in chicken breeding. The mutant site is a -734(G is greater than A) site. Genetic typing among three different groups is performed and the laying trait associated analysis is performed by means of a KASP technology, and the result shows that the site is extremely related to the laying start age and egg numbers in 32 weeks and 48 weeks (p is smaller than 0.01), and the laying start ages of genes AA and AG are extremely obviously earlier than that of GG gene, the egg numbers of the genes AA and AG in 32 weeks is extremely obviously greater than that of the gene GG but the egg number of the gene AA in 48 weeks is extremely obviously greater than those of the genes AG and GG. By detecting the molecular marker related to the laying trait, egg varieties / strains which are early to mature and high in yield are bred favorably, and favorable help is brought to local chicken breeding work.

The invention discloses an HLA (histocompatibility locus antigen) genetic typing method of an HLA determinant gene through high-throughput sequencing. Patterning HLA typing software based on various high-throughput sequencing platform data has important significance in clinic or biomedicine. Compared with the traditional sequencing method through a PCR-SBT (polymerase chain reaction-sequence based typing) method, the high-throughput sequencing technology has the obvious advantages in economic cost and time cost. HLA sequence data of thousands of samples can be read through an experiment and high resolution of HLA typing is achieved at one time through the high-throughput sequencing technology, and meanwhile, a new allele can be found. Qualitative leap in the aspects of flux detection, data quality, cost control and the like are achieved, 'low cost and high data' are achieved, additional economic burden of a patient caused by typing for multiple times can be avoided, the time for searching for a provider whose HLA is matched with that of the patient can also be reduced, and the precious time is saved for treating the patient.

The invention relates to data related to influences of different kinds of fat intake on obesity and influences of different types of sports on weight reduction and blood glucose reduction in disclosed human genome single nucleotide polymorphism (SNP) data. According to genetic differences between individuals, a method suitable for determining main reasons of obesity and making personalized diet schemes or giving suggestions about lifestyles is built, and the method is applied to development of wearable mobile devices and application software. The invention further relates to preparation and application of genetic chips related to detection. The personalized weight losing and body building schemes can reasonably, efficiently and selectively use modern sports exercise means and diet regulation according to personal gene characteristics, so that individuals achieve the aims of diet regulation and exercise planning more scientifically and more quickly.

The invention discloses a microsatellite loci marker combination which comprises 17 microsatellite loci markers, and simultaneously provides an efficient detection method based on the 17 microsatellite loci markers; the markers have high polymorphism, are not linked with each other and have proper fragments interval, and are easy to be detected simultaneously. The invention is improved in a detection technology, optimizes an experimental system by utilizing a multi-marker amplification technology and a fluorescent semi-automatic microsatellite typing method, thus establishing high flux, accurate and effective 17 cattle microsatellite marker compound amplification and a gene typing method.

The invention provides a molecular marker for a paddy recessive genic male sterility gene cyp704b2 and application thereof, belonging to the technical field of plant biology. The molecular marker provided by the invention comprises two forward primers SEQ ID NO.1 and SEQ ID NO.2 and one reverse primer SEQ ID NO.3. The molecular marker can be utilized to complete the genetic typing of the paddy recessive genic male sterility gene cyp704b2 on the basis of conventional PCR and PAGE glue electrophoresis. The molecular marker has the advantages of simplicity and convenience in operation, high typing speed, accurate result and low cost, is capable of increasing the selection efficiency of the target trait and can meet the requirements of large-scale molecular marker-assisted selection breeding.

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.

The invention discloses a method for breeding kidding traits by utilizing 3-gene pyramiding effect. The method comprises the following steps: amplifying a 3' untranslated region of a stem cell factor (SCF) gene, an exon 7 of a III type transmembrane tyrosine kinase receptor (KIT) gene and an intron 1 of a kiss (KISS1) gene respectively by using three pairs of primers and by taking goat genome DNA as a template; judging the size of each amplification product by using agarose gel electrophoresis at the concentration of 1.5%; screening the site mutation of the amplification products of the three pairs of primers by a DNA sequencing technology; performing genetic typing and gene frequency analysis on the single nucleotide polymorphisms (SNPs) of three sites of the SCF gene, the KIT gene and the KISS1 gene by using polyacrylamide gel electrophoresis at the concentration of 10% and agarose gel electrophoresis at the concentration of 3.5%; analyzing the relationship between different genotype combinations and kidding quantity; and screening out the optimal genotype combination.

The invention relates to the bio-detecting technology, in particular to a primer and a kit for detecting rice blast resistance genes Pigm and a genetic typing method. By means of the method of comparing alleles of multiple rice blast resistance genes Pigm with a 'GuMei 4' near-isogenic line sequence, a basic group difference site capable of being different from other susceptible alleles is obtained through analysis, and then design screening is carried out, so that the optimal primers S1806GC-F and S1806GC-R and optical probes S1806-VIC and S1806-FAM are obtained. The invention further discloses a genetic typing method for the rice blast resistance genes Pigm through the primers and the probes. Genetic typing analysis is carried out through the fluorescence real-time PCR technology, enzyme digestion, electrophoresis and sequencing are not needed, operation is easy and convenient, time and labor are saved, and safety and environmental friendliness are achieved.

The invention discloses an SNP marker related to rapid growth of pacific oysters and an identification method as well as application thereof, and belongs to the field of molecular biological DNA marking technology and application. By applying a high-resolution melting curve analytic technique and utilizing 108 SNP loci mined from an EST common data base to perform genetic typing on pacific oyster rapid growth population after 5 generation breeding selection, 3 alleles closely related to the growth trait of pacific oysters are obtained via correlation analysis. The 3 SNP marker preponderant alleles can be applied to selection of rapid growth parent pacific oysters, and further provide a scientific auxiliary means for breeding selection of rapid growth pacific oysters. Compared with the conventional breeding method, the method has the advantages of strong purposefulness and direct action effect, besides, the operation is simple, the detection is rapid, the cost is low, and extensive popularization and application is facilitated.

The invention belongs to the technical filed of human nucleic acid in-vitro detection, and more specifically relates to a gene typing detection kit for human 13,18 and 21 chromosome 20 STR locus. According to the invention, a composite PCR amplification system for gene typing on 20 STR locus distributed on human 13, 18 and 21 chromosome is designed, and wherein a PCR amplification primer group is SEQ ID No.1-SEQ ID No. 40. The gene typing detection kit comprises a primer combination container and a PCR reaction mother liquor container; and the primer combination container comprises primer SEQ ID No.1-SEQ ID No. 40 composition storage liquid. In the invention, single tube amplification on 20 STR locus is carried out, STR locus in each fluorescence channel and different fluorescence channels have equal amplification, a typing graph is good; cost, manpower and time can be obviously saved, and work efficiency is increased.

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.

The invention discloses primers for detecting polymorphism of CYP3A4, CYP3A5 and MDR1 genes with an ARMs-PCR (amplification refractory mutation system-polymerase chain reaction) method and a prepared kit. Primer sequences are shown in SEQ 1-SEQ 18. The primers can perform typing testing on CYP3A4, CYP3A5 and MDR1 genes of humans quickly and accurately through specific amplification after primer combination. Genetic typing or preparation of the typing kit can be performed according to the primers and the primers are used for detecting metabolic capacity of Chinese people for related drugs such as tacrolimus, cyclosporine A and the like, so that a reference basis can be provided for special doctors who determine optimal drug dosage and individual drug use, and drug use risks are reduced.

The invention provides a group-specific primer PCR-SBT reagent based on HLA-DQB1 genetic typing. The group-specific primer PCR-SBT reagent is composed of six pairs of group-specific primers for amplification and four oligonucleotides sequencing primers for sequencing analysis. The invention furthermore provides a group-specific primer PCR-SBT method based on HLA-DQB1 genetic typing adopting the reagent. The reagent and method can be used as an independent widely-applied identification method. The accurate typing identification problem of HLA-DQB1 sites can be successfully solved. Accuracy of HLA matching of hematopoietic stem cell transplantation donors and recipients can be improved. Therefore, more proper transplantation donors are selected, and rejection reactions in the transplantation process are reduced. Great significance in furthermore improving the success rate and survival rate of organ transplantation is achieved.

The invention discloses a method for keeping the genetic diversity of Jian carps. The method comprises the steps of individual tagging, template creating, genetic typing, family building, breeding file establishing and parent updating. Compared with the prior art, inbreeding of the Jian carps can be effectively avoided, the genetic diversity of Jian carp groups is kept and increased, and genetical characterization decline of the Jian carps is slowed down; in addition, breeding is assisted through molecular markers, a breeding file is established, genetic distances of the groups are analyzed by combining all parameters in the file, and therefore dominant groups are selectively reserved, and descendants excellent in genetic characterization are obtained through mating.

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.

The invention discloses a pleuroperitoneal fluid type organ culture medium and method and a drug sensitive testing method. The pleuroperitoneal fluid type organ culture medium can effective maintain specificity of tissue cells and characteristics of stem cells and ensure high consistency of genetic typing as well as high similarity of tissue shape. Besides meeting the demands of scientific research, the pleuroperitoneal fluid type organ culture medium can be applied to the aspect of clinical medication guidance, and through in vitro organ culture, provide patients with a beneficial option on medication guidance.

An oligonucleotide comprising a nucleic acid sequence selected suitable for identifying an olfactory receptor gene or an allelic variant thereof is provided.

The invention relates to a screening method of a scylla paramamosain SNPs molecular marker, which comprises the following steps of: (1) obtaining a scylla paramamosain genome sequence through a genomic library or a GenBank database obtained by sequence determination; (2) carrying out primer design on the obtained genome sequence, carrying out sequence determination after PCR (Polymerase Chain Reaction) amplification, contrasting the sequences of different individuals and verifying candidate SNPs (Single Nucleotide Polymorphisms); and (3) designing a primer with a specific allete for a candidate SNPs site and carrying out verification and genetic typing on the SNPs by adopting a real-time fluorescent quantitative PCR dissolving curve method. The screening method has the advantages of short experimental period, low cost, simplicity in operation and the like, and the genetic types of the different individuals can be accurately detected to provide a novel genetic marker for scylla paramamosain genetic variation analysis, population genetic diversity evaluation and molecular marker-assisted breeding.

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.

The invention discloses a primer, kit and method for conducting genetic typing on the hantavirus by means of a PCR direct sequencing method. The primer comprises an upstream primer: ATTAGCCCWGTCATGAGTGT, and a downstream primer: CTTTGACTCYTTTGKYTCCA, wherein the Y is a C or a T, the W is an A or a T, and the K is a G or a T. The method comprises the steps of extracting a total RNA of a virus sample to be tested, conducting reverse transcription to obtain a cDNA, using the cDNA as a template, utilizing the primer for conducting PCR amplification, conducting sequencing on an amplified target fragment, constructing a phylogenetic tree by using corresponding fragments of known genotype representative strains as a reference on the basis of sequence information of the target fragment, and conducting the genetic typing on the virus sample to be tested according to the phylogenetic tree. When the primer is used, a specificity sequence of an S gene of the hantavirus can be obtained by only conducting one-time PCR amplification, and the genetic typing can be easily, conveniently and rapidly carried out on the hantavirus by utilizing the specificity sequence.

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.

The invention discloses primers and fluorescent probes used for detecting folate metabolism related gene SNP and an application. The primers include four groups, each group having a corresponding fluorescent probe group in order to perform quick genetic typing to rs1801133, rs1801131, and rs1801394. The primers and fluorescent probes have accurate and reliable detection results and low cost, and have good practicability.

The invention relates to a primer pair for detecting CYP2C9 genetic typing through a pyrosequencing method and a kit, and belongs to the technical field of in vitro nucleic acid detection. The primer pair comprises a CYP2C9*2 forward amplification primer, a CYP2C9*2 reverse amplification primer, a CYP2C9*2 sequencing primer, a CYP2C9*3 forward amplification primer, a CYP2C9*3 reverse amplification primer and a CYP2C9*3 sequencing primer. Biotin labeling is conducted at the 5' end of the CYP2C9*2 forward amplification primer and the 5' end of the CYP2C9*3 reverse amplification primer. The kit comprises the amplification primers, a PCR reaction solution 1, a PCR reaction solution 2, the sequencing primers, uracil DNA glycosylase and Taq polymerase. The kit has the advantages of being accurate in detection result, high in specificity, short in detection period, easy to operate and capable of effectively meeting the clinical examination requirement.