48 results about "Mutant genotype" patented technology

The present invention discloses a method for breeding the commercial strains of red feather pink-shell laying hens. It provides a primer pair for identifying the red feather causative mutation homozygous genotype of white leghorn chickens, which is composed of the single-stranded DNA molecule shown in Sequence 2 of the Sequence List and the single-stranded DNA molecule shown in Sequence 3 of the list. After the primer was designed according to the upstream and downstream nucleotide sequences of the 18,288,303^rd deoxynucleotide in the positive-sense strand of the 11^th chromosome as shown in the sequence information of the chicken reference genome Gallus_gallus-4.0 version published in NCBI, the genotype (SNP) at this site is tested through the restriction fragment length polymorphism, the genotype of the site (SNP) was tested through the restriction fragment length polymorphism; the offspring hens obtained by cross breeding the homogenous female parent (the homogenous female parent was obtained through expanded propagation of the white leghorn chickens with the red feather causative mutation homozygous genotype) and the Rhode Island Red rooster as a male parent are all of red feather phenotype, meeting the market demands and enjoying a broad prospect for promotion.

The invention belongs to the technical field of genome sequencing, and discloses an insertion mutation detection method based on new-generation sequencing data. The method comprises the steps: when amutation generation site is determined, a region where insertion mutation occurs certainly generates a split reading section, aiming at the characteristics that insertion variation types such as new sequence insertion, sequence series multiplication and sequence dispersion multiplication are different in distribution of missing variation and inverted mutation split reading sections, constructing avirtual reference sequence by utilizing partial matching, complete matching and unmatched read segment information after determining the insertion mutation generation type and site, and comparing thevirtual reference sequence with the original reference sequence to obtain related information of the insertion sequence; and obtaining a mutant genotype by utilizing the copy number state information. According to the invention, the problem of inaccurate insertion mutation point judgment can be solved; the problem of omission caused by insertion mutation detection of an SR method can be solved; the problem that in the prior art, repeated sequences may cause detection errors can be solved.

The invention discloses a PIK3CA gene mutation fluorescence quantitative PCR genotype detection kit comprising a PCR mixed reaction liquid, a positive control, and a fluorescent probe used for detecting PIK3CA gene mutant genotypes. The PCR mixed reaction liquid comprises PCR primers used for amplifying PIK3CA gene segments where the mutation sites exist. The invention also discloses a PIK3CA gene mutation fluorescence quantitative PCR genotype detection method. According to the method, PIK3CA gene mutant genotypes are subjected to fluorescence quantitative PCR detections by using the kit provided by the invention. With the technical scheme provided by the invention, detections can be carried out upon tissue PIK3CA gene mutations, and especially rapid, accurate, and high-sensitively detections upon PIK3CA gene 542,545 and 1047 nucleotide mutations can be realized.

The invention relates to a special CAPS marker for identifying a wild type or mutant paddy rice salt resistant gene OSRR22, a primer, and primer applications. The marker comprises a marker Cas-Osrr22-1 for detecting a wild type site and the nucleotide sequence of the marker Cas-Osrr22-1 is represented by SEQ ID No.2. The marker also comprises a marker Cas-Osrr-22-2 for detecting a mutant WDR58-cas-1 site, and the nucleotide sequence of the marker Cas-Osrr-22-2 is represented by SEQ ID No.3. Compared with the prior art, the provided maker, primer, and primer applications have the advantages that a molecular marker for differentiating mutant and wild type of WDR58-cas-1 is provided, high efficient identification of salt resistant gene Osrr22 mutant genotype is realized; and WDR58-cas-1 mutant can be applied to breeding of salt resistant species. The CAPS marker can be applied to assistant breeding for improving the salt resistant property of paddy rice, and can be applied to identification for crossbreeding and backcross separation groups, heterozygosis genotype and homozygosis genotype can be accurately differentiated, and the breeding efficiency is improved.

The invention discloses a red feather mutant of white leghorn as well as a marker and a detection method of the red feather mutant. The invention provides application of a substance for detecting thegenotype of red feather SNP (Single Nucleotide Polymorphism) luci in a genome of the white leghorn in identification of color characters of feather of filial generations of to-be-detected white leghorns and Rhode Island Red cocks; the red feather SNP loci are the 18th,, 288th and 303rd loca of a positive-sense strand of No.11 chromosome of a chicken genome. The experiment of the invention proves that one SNP locus is found and can realize identification of the red feather mutagenic genotype; by using the method, the red feather mutagenic genotype of white leghorn strain chicken is successfullyidentified. The identification method is convenient and quick, can be used for quickly cultivating a novel red feather mutant homozygotic type strain and further is applied to cultivation of a novelcomplete strain of 100 percent red feather phenotype red feather pink egg-shell chickens; cultivation cost is reduced, cultivation time is shortened and variety benefits are increased.

A muscovy duck egg laying trait associated gene SMAD3 molecular marker screening method comprises the following steps: (1) carrying out DNA extraction: extracting female muscovy duck genome DNA in anegg laying period by adopting a phenol-chlorine method, and then determining an OD value and concentration of the purity of the genome DNA; (2) carrying out primer design: carrying out SMAD3 gene amplification primer design by applying NCBI online software and Primer 5.0, and sending a primer sequence to a related biological company for synthesis; (3) carrying out PCR amplification: adopting a 15[mu]L reaction system, setting PCR amplification conditions, carrying out PCR amplification, and carrying out electrophoresis detection on a product by using 1% agarose gel; (4) sequencing amplification product fragments and screening SNP sites; and (5) directly sequencing and typing the SNP mutation sites, carrying out correlation analysis on the SNP mutation sites and the egg laying performance,and determining the SNP closely correlated with the egg laying performance. When the molecular marker is applied to the muscovy duck seed selection and breeding process, the mutant genotype of the gene SMAD3 associated with the muscovy duck egg laying performance detected by the method is used as the molecular marker.

The invention belongs to the technical field of biological detection and discloses a kit for accurately detecting the polymorphism of an ALDH2 (Aldehyde Dehydrogenase 2) gene. The kit for accurately detecting the polymorphism of the ALDH2 gene can be used for determining mutation gene types (G>A, G>T and G>C) of rs671 polymorphism sites of the ALDH2 gene in one step. In order to realize the function, the kit applies a method combining a multi-PCR (Polymerase Chain Reaction) technology and a Taqman-MGB fluorescent probe labeling technology, and four pairs of specific primers and four specific probes for labeling different fluorescent dyestuffs are designed; the kit is characterized in that a target gene only can be combined with one pair of primers and one probe, so that the accuracy of a detection result is ensured. The kit provided by the invention has the advantages of strong specificity, high accuracy, simplicity in operation and the like. A sample to be detected can be selected from whole blood or oral cells; the kit is clinically used for guiding rational drug use of nitroglycerin and guiding healthy wine drinking to prevent alcohol intoxication, and has great significance inthe aspect of prompting major diseases.

The invention discloses a method for detecting a genotype of polymorphic sites of a corneal dystrophy gene and its kit. In a same reaction system, real-time fluorescence quantification PCR detection is respectively carried out on a same to-be-detected sample by an enhanced ARMS primer aiming at a gene wild type template for polymorphic sites of corneal dystrophy and an enhanced ARMS primer aimingat a mutant template; according to a Ct mutant type detected by the real-time fluorescence quantification detection by the enhanced ARMS primer aiming at the gene wild type template, a Ct wild type detected by the real-time fluorescence quantification detection by the enhanced ARMS primer aiming at the mutant template, and a difference delta-Ct value of the Ct mutant type and the Ct wild type, and the mutant genotype of the corneal dystrophy gene can be determined according to the types of the polymorphic site. The method has the advantages of simple operation and high accuracy.

The invention relates to the technical field of bioinformatics, in particular to a method for screening disease phenotype related mutation sites and application thereof. The method comprises the following steps: obtaining sequencing data of a plurality of disease samples and normal samples, and carrying out variation detection; performing association rule mining by taking the phenotype of the samples and the mutation type of the detected mutation site as a total project set to obtain a mutation site having a strong association relationship with the phenotype of the disease samples; and performing modeling analysis on the mutation sites obtained through association rule mining and screening to obtain mutation sites related to disease phenotypes. Alleles are converted into classification variables for association rule mining, and then modeling analysis is carried out on sites strongly associated with disease phenotypes, so that the total quantity of analyzed samples can be effectively reduced, and the influence of allele frequency on an analysis result is avoided; and screening and analysis of disease phenotype related sites can be completed only by obtaining mutation genotype information.

The invention relates to a method for identifying plant gene functions in total genome ranges.The method includes 1), mutating single nucleotide of starting plants and establishing plant mutant libraries; 2), detecting phenotypes of mutants in the plant mutant libraries and establishing plant mutant phenotype databases; 3), detecting genotypes of the mutants in the plant mutant libraries and establishing plant mutant genotype databases; 4), determining functions of genes where SNP (single nucleotide polymorphism) sites caused by mutation of the single nucleotide in the starting plants by the aid of the plant mutant phenotype databases and the plant mutant genotype databases.The method has the advantages that the genes related to plant development procedures and biological procedures of stress-tolerant reaction or metabolic pathways or the like can be accurately and quickly found by the aid of the plant mutant phenotype databases and the plant mutant genotype databases which are established by the method, and novel ways can be created for plant functional genome research.

Recessive male sterility has an important utilization value in plant breeding. TDR gene function deficits in rice can cause thorough recessive male sterility, and is thus an important target of creating the recessive male sterility in genetic engineering. The invention discloses a method for performing site-directed mutation on a rice TDR gene by a CRISPR\Cas9 system. By using the site-directed mutation method provided by the invention, a TDR gene mutant can be fast and efficiently created so as to obtain a recessive male sterility material. The invention further provides a detection method ona specific TDR mutation genotype obtained by the site-directed mutation method. The detection method can be used for fast identifying whether the mutation exists in the target in a current generationtransformant of the TDR site-directed mutation, and can also be used for genotyping identification of the later-generation specific mutation genotype.

High-throughput detection for the interesting base or the mutation site in the nucleic acid sample can be achieved by the linear test probe pairs P1 and P2. The test probe pairs P1 and P2 respectively comprise either of the flanking complementary sequences which are adjacent to the interesting base or the mutation site in the nucleic acid sample. When the test probe pairs P1, P2 are annealed and hybridized to the nucleic acid sample, a gap will be generated at the interesting base or the mutation site position between the probe pairs and the sample. Divide the annealed hybrid sample into four equal reaction systems to which add dATP, dTTP, dCTP, dGTP, respectively. The test probe pairs P1 and P2 will be ligated into one single probe when adding the complementary nucleotide system under the DNA polymerase or ligase. After purified and amplified, the generated single probes are hybridized to the corresponding area in a common oligonucleotide microarray. The generated single probe will give a signal in the hybrid area, and therefore detect and analyze the hybrid signal to determine the base type or the mutation genotype at the detection position. The invention can be applied to the re-sequencing the target nucleic acid sequence, the detection and analysis for the mutation, insertion, or deletion sites of a known nucleic acid sequence, and the genotyping of the pathogenic microorganism.

The invention discloses a screening method of a molecular marker of a circadian rhythm behavior related gene Cry1 of egg ducks. The screening method comprises the following steps: (1) extraction of DNA: extracting DNA by using a blood/cell/tissue genome DNA kit, and then assaying the OD value and concentration; (2) designing of a primer: designing the primer by using Primer Primer 5.0, and delivering the primer to a biology company for synthesis; (3) PCR amplification: conducting PCR amplification by using a 20mu L reaction system and setting the PCR amplification conditions, and taking and detecting the product by agarose gel electrophoresis; (4) fragment purification of the amplification product, vector linking, conversion, positive clone screening and sequencing; and (5) SNP mutant site screening and correlation analysis for determining the circadian rhythm behavior related SNP of egg ducks. In the molecular breeding process, the mutant gene type of the circadian rhythm behavior related gene Cry1 detected by the method is regarded as the molecular marker.

The invention discloses a method for guiding an editing system to mediate crops to generate endogenous herbicide resistance. According to the invention, various herbicide-resistant ACCase mutant genotypes are obtained, and the ACCase mutant genes respectively obtain single amino acid mutation at different sites. The ACCase mutant gene is obtained by utilizing a CRISPR-mediated guide editing system, specifically, the 1879th amino acid is mutated into leucine, threonine, valine, serine, methionine and the like from isoleucine, the 1927th amino acid is mutated into tyrosine or phenylalanine from proline, the 2097th amino acid is mutated into glycine, serine or leucine from tryptophan, and the ACCase mutant gene is mutated into leucine, threonine, valine, serine, methionine and the like from tryptophan. The 2139th site is mutated into aspartic acid, valine and serine from isoleucine, the 2176th site is mutated into glycine from aspartic acid, and the 2194th site is mutated into serine and alanine from glycine. These ACCase mutant genes endow receptor plants with resistance (tolerance) to acetyl-CoA carboxylase inhibitor herbicides to varying degrees.

The invention belongs to the technical field of animal breeding test and particularly discloses a method for detecting slow-feathering mutation genotypes of cocks. The method includes the following steps of S1, extracting DNA (deoxyribonucleic acid) of blood samples of detected individuals and diluting the DNA to be 40-100nmol/mu L; S2, performing real-time fluorescence quantitative PCR (polymerase chain reaction), wherein the type of a fluorescence quantitative PCR instrument is United States Bio-rad CFX96, a fluorescence quantitative reagent is Bio-rad SYBR Green fluorescence quantitative PCR Mix, a fluorescence quantitative primer sequence is as shown in SEQ ID NO: 1-4; S3, calculating delta Ct value of the detected individuals, and judging the genotypes of the individuals according to the delta Ct, where when the delta Ct is greater than 1.5, the genotypes are heterozygotes, when 0.3 is less than delta Ct which is less than 1.3, the genotypes are homozygotes, and when 1.5 is greater than or equal to delta Ct which is greater than or equal to 1.3, the genotypes are eliminated. The method has the advantages of short period, high speed, 100% of accuracy, less steps, less proneness to making mistakes and the like.