38 results about "Single nucleotide mutation" patented technology

The invention discloses a single nucleotide mutation site SNP (Single Nucleotide Polymorphism) and KASP (Kinase Associated Specific Protein) marker remarkably associated with soybean plant height and application thereof. The SNP molecular marker is located at the 36108741bp position of the fifth chromosome of the soybean, the basic group is T or C, the SNP molecular marker is significantly related to the plant height phenotype of the soybean, and the plant height of the soybean variety with the locus genotype of TT is significantly lower than that of the soybean variety with the genotype of CC; three KASP primers are developed according to the SNP site and are respectively SEQ ID NO.1, SEQ ID NO.2 and SEQ ID NO.3, PCR amplification and genetic typing are carried out on to-be-detected soybeans by utilizing the primers, and if a detection result shows that the basic group type at the marker position is T, it is judged that the plant height of the soybean variety is relatively low; if the detection result is C, it is judged that the plant height is high. The SNP molecular marker disclosed by the invention can be used as an auxiliary selection marker for the plant height character in the soybean breeding process, so that the selection accuracy is improved, and the soybean plant height character related breeding process is accelerated.

The invention discloses an SNP (Single Nucleotide Polymorphism) marker related to a soybean branching number gene QNE1 and application thereof, and relates to the technical field of molecular genetics and crop genetic breeding, the SNP molecular marker is a nucleotide sequence as shown in SEQ ID NO.1 formed by single nucleotide mutation at the 686th site of the nucleotide sequence of the QNE1 gene in a soybean genome; the polymorphism of the single nucleotide mutation is C / G polymorphism. The method is used for assisting soybean breeding. The SNP marker provided by the invention can be used for molecular marker-assisted selection of soybean branch number related traits, and a key tool is provided for cultivating soybean varieties adapting to different planting requirements (such as a few-branch close planting type or a multi-branch high-yield type).

The present application relates to the technical field of tomato genetic breeding, and more specifically to molecular markers associated with tomato traits under low-phosphorus stress and their applications. The molecular marker comprises a nucleotide sequence formed by a single nucleotide A>G mutation at position 52526904 of chromosome 11 of the tomato genome. Using this molecular marker, not only can the plant height, fresh weight, and dry weight of tomatoes at the seedling stage under low-phosphorus stress be predicted and identified, but the degree of low-phosphorus tolerance of the plant can also be determined. This molecular marker can assist in cloning genes controlling plant height, fresh weight, and dry weight of tomatoes at the seedling stage under low-phosphorus conditions and in breeding tomatoes for low-phosphorus tolerance, and has important application value.

The application discloses a single cell whole genome amplification sequencing method. The application comprises the following steps: 1) separating a single cell, adding a single cell lysis solution to sufficiently lyse the single cell and digest proteins combined on the genomic DNA; 2) using a Tn5 transposome with a specific adapter sequence to fragment the DNA, while adding adapters to both ends of the fragments; 3) after end repair, using a ribonuclease to cut modified ribonucleotide residues on the adapter sequence; 4) using the adapter sequence as a primer to linearly amplify the fragmented DNA by using a single primer PCR method; and 5) adding sequencing adapters to both ends of the DNA fragments by using a chain extension and PCR method to obtain a final sequencing library. The method disclosed by the application can amplify single cell whole genome DNA with high coverage, uniformity and fidelity, and simultaneously and accurately detect chromosomal copy number variation CNV and single nucleotide mutation SNV on a single cell genome.

The application provides a global ancestor estimation method and system based on principal component analysis, which comprises the following steps: based on the principal component analysis method, according to single nucleotide mutation data, projecting the principal components of the to-be-tested offspring samples and the principal components of the ancestral samples into the same two-dimensional plane, and dividing the projected two-dimensional plane into multiple corresponding population regions; fitting the population region of each ancestral sample to obtain an ancestral population fitting region, and obtaining the region density of each ancestral population fitting region; extending each ancestral population fitting region outward at the same proportion, stopping the outward extension when any ancestral population fitting region contains a to-be-tested offspring sample, and obtaining the ancestral population proportion of the current to-be-tested offspring sample according to the region area after the extension and the region density. The application directly calculates the proportion of the corresponding ancestor in the offspring individual or population through the principal component analysis result, and has high accuracy.

The invention relates to a product for KRAS gene detection based on combination of a dumbbell-shaped probe and RCA, the product comprises the dumbbell-shaped probe, DNA ligase and an RCA amplification reagent, the dumbbell-shaped probe comprises a stem part and a two-ring part, the ends of the two-ring part are connected through the stem part, one ring of the dumbbell-shaped probe contains a G-quadruplex aptamer PW17 complementary sequence, and the other ring of the dumbbell-shaped probe contains a G-quadruplex aptamer PW17 complementary sequence. One ring contains a complementary sequence of a G-quadruplex aptamer T30695, the other ring contains a complementary sequence of a G-quadruplex aptamer T30695, the RCA amplification reagent comprises Phi29DNA polymerase, dNTPs and a Phi29DNA polymerase reaction buffer solution, the stem part of the dumbbell-shaped probe is opened under the mediation of DNA ligase and a detection gene, and the cut is connected to form a closed ring structure which is used as a template of the RCA. By adopting the technical scheme, the invention provides a product for detecting the KRAS gene based on the combination of the dumbbell probe and the RCA, and compared with the traditional method, the product is more convenient and more efficient when the mononucleotide mutation of the gene is tested.

The invention relates to the technical field of plant molecular breeding and molecular biology, in particular to a KASP molecular marker related to soybean mosaic virus resistance and application of the KASP molecular marker. The molecular marker is targeted to an anti-soybean mosaic virus specific functional SNP (Single Nucleotide Polymorphism) site of a soybean GmLMM4 gene; the SNP locus is c.743Cgt of a soybean GmLMM4 gene coding region, and the SNP locus is a nucleotide sequence of the soybean GmLMM4 gene; the GmLMM4 gene is mutated by a T mononucleotide, and the 248th amino acid of the GmLMM4 gene coding protein is changed into isoleucine from threonine through the mutation. The molecular marking tool provided by the invention has the advantages that the accuracy of anti-SMV breeding can be remarkably improved, the matched primer group is designed in a targeted antisense strand, and the C / C, C / T and T / T genotypes can be accurately distinguished by FAM / HEX fluorescence signals and correspond to soybean anti-SMV phenotypes; high-throughput detection is completed in the seedling stage, the screening efficiency of high-resistance materials is improved, the identification period is shortened, and the screening efficiency of soybean anti-SMV breeding is improved.

This invention relates to a GmSPL3b gene SNP marker and its application, belonging to the fields of plant molecular biology and crop genetics and breeding technology. The SNP marker is used for molecular detection of soybean flowering and maturity stages. The SNP is a single nucleotide mutation at position 94 and / or position 116 of the first exon of the soybean GmSPL3b gene, forming a nucleotide sequence as shown in SEQ ID NO. 1 and / or SEQ ID NO. 2; the polymorphism of the single nucleotide mutation is A / G polymorphism. This invention enables precise genotyping at the seedling stage, efficiently breeding early-maturing soybean varieties adapted to high-latitude regions. It can directly serve as a precise target for gene editing, providing a clear basis for molecular design breeding. It allows for rapid screening of ideal genotypes in early generations, reducing field workload, shortening the breeding cycle, and accelerating the selection of new varieties.

Methods and materials are provided for detecting nucleic acid sequence differences including single nucleotide mutations or polymorphisms, one or more nucleotide insertions, and one or more nucleotide deletions in single molecule target members present in a test population of nucleic acid fragments. Heteroduplexes are formed between members of the test nucleic acid population and their corresponding complements provided in a pool of mismatch cleavage probes. Mismatched base pairs in the heteroduplexes are specifically cleaved and cleaved probe fragments are electronically detected to signal the present of the target members in the test population.

The application relates to a mutant type of an INSIG1 gene for regulating fat deposition of animals and application thereof, wherein the mutation is located at the 3725th base of a nucleotide sequence shown in SEQ ID NO. 1, and is an A>G single nucleotide mutation. The application analyzes the genetic basis of fat deposition of animals by using comparative genomics, finds that a strictly conserved site of an INSIG1 gene corresponds to parallel replacement of histidine to arginine of amino acids, and the potential function of the strictly conserved site in fat deposition and mobilization is prompted.

The present disclosure relates to an apparatus and a method for providing information for diagnosing micro-residual diseases through quantification of single-nucleotide variants in cfDNA. The apparatus according to the present disclosure comprises: a memory in which at least one process for performing an operation for providing information for diagnosing micro-residual diseases is stored; and a processor for performing an operation according to the process, wherein the processor analyzes WGS of patient-derived cancer tissues using two or more pieces of somatic variant analysis software to obtain data on the positions of consensus single-nucleotide variants commonly identified in primary cancer samples, matches reference sequences at the positions of the single-nucleotide variants with cfDNA WGS of the patient to obtain supporting reads matching in which the variants match in overlapping regions of paired reads, filters the obtained supporting reads on the basis of predetermined conditions, and generates probability data related to micro-residual diseases on the basis of the filtered supporting reads.

The invention belongs to the technical field of molecular markers, and particularly relates to a KASP molecular marker related to wheat agronomic traits and application of the KASP molecular marker. The nucleotide sequence of the KASP molecular marker is shown as SEQ ID NO.2, and single nucleotide T-G mutation occurs at the 101bp site. The molecular marker is a KASP molecular marker developed by aiming at an SNP (Single Nucleotide Polymorphism) site of a TaFLA62 promoter region of a bundled arabinogalactoprotein-like gene. The mutation from T to G at the 101bp of the KASP molecular marker can reduce the wheat plant height and / or increase the grain weight. The molecular marker provided by the invention can efficiently detect and track a mutation site of a TaFLA62 gene promoter region in a wheat variety / strain, and provides technical support for wheat molecular breeding.

This application relates to molecular markers associated with pork intramuscular fat and their applications. The SNP is located at a single nucleotide polymorphism (G>A) at bp 6114468 on chromosome 1 of the porcine genome. PCR amplification of the target sequence containing the single nucleotide mutation site using primers and sequencing of the amplified product can rapidly identify PRKN genotypes, thereby detecting differences in pork quality and providing guidance for pig breed selection.

The application relates to a WEE1 gene molecular marker related to pig carcass oblique length and application thereof, relates to the technical field of molecular genetic markers, and aims to provide a SNP molecular marker related to pig carcass oblique length and application thereof, and to apply the same in genotype identification, genetic breeding and the like, and aims to screen a molecular marker related to pig carcass oblique length. The SNP molecular marker of the application is a nucleotide sequence shown as SEQ ID NO. 1 formed by single nucleotide mutation of the 616th base of the WEE1 gene; the polymorphism of the single nucleotide mutation is T / G polymorphism. The application is used for breeding / assisting breeding of pig varieties or strains related to pig carcass oblique length. The application identifies the pig carcass oblique length by judging the polymorphism of the mutation site of the pig WEE1 gene promoter region, and applies the mutation site as early selection, so as to provide a new idea for improving the pork yield of pigs. The application has important breeding significance for improving the pork yield of pigs.

The invention relates to a method for improving soybean alkali stress resistance based on an autophagy key gene and application of the method, and belongs to the technical field of plant genetic engineering and crop genetic breeding. An autophagy related gene GmATG8c is overexpressed in the soybean and is combined with specific mononucleotide mutation located on a 20th chromosome, so that the alkali resistance of the soybean is synergistically enhanced. Experiments prove that the germination rate, the plant survival rate and the biomass of a transgenic soybean strain (NK-Al) simultaneously carrying GmATG8c gene overexpression and mononucleotide mutation under an alkali stress condition are obviously superior to those of a wild type strain, a strain independently carrying GmATG8c overexpression and a strain independently carrying the mutation. The invention discloses a new mechanism for synergistically regulating and controlling the alkali resistance of the plant by a cell autophagy pathway and a specific transcription factor, and provides a new gene resource and a technical strategy for cultivating alkali-resistant soybeans and other new important crop varieties.

The present invention belongs to the field of genetic breeding technology and specifically relates to a molecular marker for breeding lean-meat sheep and its application. The nucleotide sequence of the molecular marker is shown in SEQ ID NO. 1, and a single nucleotide mutation from C to T occurs at the 101st bp position of the molecular marker. The present invention provides a SNP locus and related molecular markers that affect the fat thickness ratio of sheep. By selecting individuals with the CC genotype as sires or dams, the fat thickness ratio of offspring can be reduced.

The invention belongs to the technical field of animal genetic breeding, and provides application of an SNP molecular marker associated with a pig backfat thickness character. The nucleotide sequence of the molecular marker is as shown in SEQ ID NO: 7, wherein the polymorphism of the 88th basic group is T / C. The method comprises the following steps: performing deep sequencing on individual genomes with different backfat thicknesses of large white pigs, and finding that mononucleotide mutation Tgt exists at the 88th basic group of the 15th exon of the GLB1 gene; and C, inter-group difference exists. According to the present invention, the pig population is further expanded, the genome DNA is subjected to PCR amplification sequencing, the mutation site typing is performed, the correlation analysis with the corrected 100 kg backfat thickness of the large white pig is performed, and the correlation with the pig backfat thickness is determined, such that the reference data can be provided for the molecular marker-assisted breeding of the large white pig, and the continuous requirement of the large-scale production on the efficient feed energy utilization can be easily met.

The invention belongs to the field of biomedical engineering, relates to a human mitochondrial genome detection technology, and provides a human mitochondrial gene mononucleotide mutation detection method which comprises the following steps: extracting whole genome DNA of a sample; carrying out UM I labeling and purification on the mtDNA; carrying out targeted amplification enrichment and purification on the mtDNA connected with the UMI; constructing an mtDNA library; and performing computer sequencing by using a nanopore sequencing technology to obtain sequencing data, and analyzing the sequencing data to obtain a single gene mutation result. According to the method disclosed by the invention, by virtue of UM I library building and Nanopore sequencing technologies, the situation that obtained Nanopore sequencing data contains sequences except mtDNA, such as nuclear DNA, can be avoided, so that an mtDNA mononucleotide mutation result obtained after analysis of the Nanopore sequencing data is not distorted and is closer to a real situation.

The invention discloses application of an HSPB7 gene molecular marker related to pig skeletal muscle development, relates to the technical field of molecular genetic markers, and particularly relates to application of the HSPB7 gene molecular marker in breeding / assisted breeding of pig varieties or strains with pig skeletal muscle development. The HSPB7 gene molecular marker is a nucleotide sequence as shown in SEQ ID NO.1, which is formed by single nucleotide mutation of a 3'UTR (Untranslated Region) region + 54-site basic group of an HSPB7 gene; the polymorphism of the single nucleotide mutation is shown as G / T polymorphism. Experiments show that the activity of the HSPB7 3 'UTR region is remarkably improved by T-G mutation of the + 54 site of the 3' UTR region of the pig HSPB7 gene in an in-vitro test, and the difference between the T-G mutation and the activity of the HSPB7 3 'UTR region is remarkable, so that the method for detecting the molecular marker associated with the mutation is simple, convenient and rapid, is not influenced by the environment, and can realize early-stage seed selection.

The application discloses a single nucleotide mutation site SNP significantly related to soybean hundred-grain weight, a KASP marker and application thereof. The SNP molecular marker is located at the position of 40068386bp of the 15th chromosome of soybean, and is significantly related to the hundred-grain weight phenotype of soybean. The hundred-grain weight of the soybean variety with the genotype TT is significantly lower than that of the soybean variety with the genotype CC. Three KASP primers are developed according to the SNP site, and are respectively SEQ ID NO. 1, SEQ ID NO. 2 and SEQ ID NO. 3. The primers are used for PCR amplification and genotyping of the soybean to be tested. If the detection result shows that the base type at the marker position is T, it is determined that the hundred-grain weight of the soybean variety is small. If the detection result is C, it is determined that the hundred-grain weight is large. The SNP molecular marker can be used as an auxiliary selection marker of the hundred-grain weight trait in the breeding process of soybean, improves the selection accuracy, and speeds up the breeding process related to the hundred-grain weight trait of soybean.

The application discloses a single nucleotide mutation site SNP significantly related to soybean plant height, a KASP marker and application thereof. The SNP molecular marker is located at the position of 36108741 bp of the 5th chromosome of soybean, and is significantly related to the soybean plant height phenotype, and the plant height of the soybean variety with the TT genotype is significantly lower than that of the soybean variety with the CC genotype. Three KASP primers are developed according to the SNP site, and are respectively SEQ ID NO. 1, SEQ ID NO. 2 and SEQ ID NO. 3. The primers are used for PCR amplification and genotyping of the soybean to be tested. If the detection result shows that the base type at the marker position is T, it is determined that the soybean variety has a short plant height. If the detection result is C, it is determined that the plant height is high. The SNP molecular marker can be used as an auxiliary selection marker of the plant height trait in the soybean breeding process, improves the selection accuracy, and speeds up the breeding process related to the soybean plant height trait.

The application provides a model for identifying allele imbalance markers driving tumor evolution based on a hierarchical Bayesian framework and a construction method thereof, and belongs to the field of information technology.The application provides a construction method of a model for identifying allele imbalance markers driving tumor evolution based on a hierarchical Bayesian framework, a three-level Bayesian hierarchical model is constructed, global noise, subclone specificity, regional discreteness and allele single nucleotide mutation (SNV) site observation are jointly modeled, and a reparameterization correction allele copy number deviation is introduced.On this basis, a Markov Monte Carlo (MCMC) sampling is used to obtain a posterior distribution of parameters, and a sample comparison method of a posterior distribution probability of each parameter including a true allele imbalance coefficient and a Kullback-Leibler divergence is provided, which can be used for identifying allele imbalance and analyzing markers driving evolution.

This application relates to a molecular marker, primer, kit, method and application in the sixth intron of the porcine PRKN gene. The molecular marker includes a nucleotide sequence formed by a single nucleotide A>C mutation at position 6,447,587 on chromosome 1 of the porcine genome. Using a primer pair to perform PCR amplification on a target sequence containing this single nucleotide mutation site, sequencing the amplification product can quickly identify the PRKN genotype, thereby detecting differences in pork quality and providing guidance for the breeding of pig breeds.

The present application relates to a product based on dumbbell-shaped probe and RCA for KRAS gene detection, the product comprises a dumbbell-shaped probe, a DNA ligase and RCA amplification reagents, the dumbbell-shaped probe comprises a stem and two loop parts, the ends of the two loop parts are connected through the stem, one of the loops of the dumbbell-shaped probe contains a G-quadruplex aptamer PW17 complementary sequence, and the other loop contains a G-quadruplex aptamer T30695 complementary sequence, the RCA amplification reagents comprise Phi29 DNA polymerase, dNTPs and Phi29 DNA polymerase reaction buffer, under the mediation of the DNA ligase and the detected gene, the stem of the dumbbell-shaped probe is opened, the nick ligation forms a closed loop structure, and serves as a template for RCA. By adopting the technical scheme, the present application provides a product based on dumbbell-shaped probe and RCA for KRAS gene detection, compared with the traditional method, when testing single nucleotide mutation of the gene, it is more convenient and more efficient.

The application relates to a molecular marker, a primer, a kit, a method and application in a third intron of a pig PRKN gene. The molecular marker comprises a nucleotide sequence formed by a single nucleotide G>A mutation at 5964857 bp of a first chromosome of a pig genome. The primer is used for PCR amplification of a target sequence containing the single nucleotide mutation site, and sequencing of the amplification product can quickly identify the PRKN genotype, thereby detecting the difference of pork quality and providing guidance for pig breeding.

The invention discloses a sex identification method for seriola aureovittata, which comprises the following steps: (1) designing a primer for amplifying a DNA (Deoxyribonucleic Acid) fragment according to the sex-related DNA fragment of the seriola aureovittata; (2) extracting DNA (deoxyribonucleic acid) of a seriola quinqueradiata individual to be subjected to sex identification, performing PCR (polymerase chain reaction) by taking the DNA as a template and adopting the primer in the step (1), obtaining a PCR amplification product of the seriola quinqueradiata individual to be subjected to sex identification after the reaction is finished, performing enzyme digestion on the PCR amplification product, and performing electrophoresis identification on the product after the enzyme digestion; according to the invention, one mononucleotide mutation site GAT (T / C) ATC between the sexual seriola quinqueradiata male and female individuals is utilized to construct an enzyme digestion map capable of correctly distinguishing the sexual seriola quinqueradiata male and female individuals, so that the PCR-RFLP method for accurately and rapidly identifying sexual seriola quinqueradiata is established.

The invention relates to the technical field of biological detection, in particular to a single nucleotide mutation detection probe and method integrating a dynamic signal and an accumulated signal. The single nucleotide mutation detection probe comprises a DNA hairpin H1, an RNA base embedded hairpin Sub, a DNA single strand E1, a DNA single strand E2 and a DNA hairpin H2, wherein the DNA hairpin H1, the RNA base embedded hairpin Sub, the DNA single strand E1 and the DNA single strand E2 form a DNA enzyme structure; the 5'terminal of the DNA hairpin H2 is modified with carboxyl molecules, and the 3 'terminal of the DNA hairpin H2 is modified with ferrocene molecules. The hairpin probe recognition technology is combined with DNA enzyme, the purpose of cyclic detection is achieved through the thermodynamic chain competition effect, the sensitivity of single base mutation detection is improved through an electrogenerated chemiluminescence detection method for indirectly detecting a mononucleotide mutant by recognizing an intermediate product chain in a cyclic reaction, and the detection sensitivity of the mononucleotide mutant is improved. And the accuracy of single base mutation recognition is enhanced.

The invention discloses an SNP (Single Nucleotide Polymorphism) molecular marker associated with stress-induced tomato fallen leaves and application. The SNP molecular marker comprises a 57663293th basic group Ggt of a No.3 chromosome of a tomato genome; the nucleotide sequence is formed by mutation of T mononucleotide. Through the SNP molecular marker, the related defects in a conventional breeding method can be effectively overcome, including that fallen leaves cannot be judged in an unstressed state, and the judgment of the traits of the fallen leaves is influenced by the environment; the SNP molecular marker can be used for accurately and efficiently identifying the phenotype of fallen leaves under stress in the tomato seedling stage, so that the planting scale of breeding materials is effectively reduced, the workload of phenotype identification in the later period is reduced, excellent target single plants are obtained through rapid screening, the breeding process is accelerated, and the breeding efficiency is improved.

The present application relates to the technical field of tomato genetic breeding, and specifically to molecular markers associated with ammonium tolerance in tomato seedlings and their applications. The molecular marker comprises a nucleotide sequence formed by a single nucleotide A>G mutation at position 62760934 of chromosome 8 of the tomato genome SL2.50ITAG2.4. The molecular markers, nucleic acid molecules, primer pairs, kits, methods and applications provided in the present application can accurately and efficiently determine the ammonium tolerance of tomatoes at the seedling stage, effectively reducing the planting scale of breeding materials, alleviating the workload of later field phenotypic identification, accelerating the breeding process and improving breeding efficiency. The molecular marker is a universal molecular marker with little restriction on genetic materials and a wide range of applications.

This embodiment of the specification discloses molecular markers related to rice tiller number regulation genes and their applications. The molecular markers include single nucleotide mutations at positions 28,427,378 bp and 28,513,517 bp on chromosome 3 of the rice genome, and two nucleotide insertions at positions 28,428,063 bp and 28,428,347 bp. The molecular markers are respectively: SNP_3_28427378_A / G, INDEL_3_28428063_C / CAT, INDEL_3_28428347_T / TAC, SNP_3_28513517_C / T. Using the molecular markers provided by the present invention, genotypes with more tiller numbers of rice OsTB1 and OsSLR1 genes can be detected quickly and accurately, which is beneficial to the efficient application of these genes in improving the plant type structure of rice.