193 results about "Reference genes" patented technology

A molecular marker closely linked to a pepper fruit shape gene. Taking a basic version of Zhangshugang pepper as a reference gene, a single nucleotide polymorphism is present at base 162028839 of the pepper Chr02 chromosome, where a substitution from base C to A occurs. A primer and kit for identifying the molecular marker closely linked to the pepper fruit shape gene, and use thereof in identifying the pepper fruit shape or in molecular-assisted breeding. Early use of the molecular marker, the primer, and the kit in the early stage can quickly screen satisfactory plants, thereby effectively reducing the planting scale, reducing the workload of later identification, and improving the selection efficiency and accuracy. The molecular marker, the primer, and the kit are of great significance in the practice of pepper fruit shape breeding and the research on fruit shape changes and regulation mechanisms.

The invention relates to the technical field of molecular biology, in particular to a fluorescent quantitative PCR reference gene screening method in different tissues and stress treatment of perilla frutescens, which specifically comprises the following steps: step 1, respectively selecting perilla frutescens leaves, stalks, roots, fruits at different development stages after flowering and perilla frutescens leaves treated by 20% PEG6000 for 0H, 3H, 6H, 9H, 12H and 24H as samples; extracting total RNA (Ribonucleic Acid) of each sample, and carrying out reverse transcription on the RNA to synthesize cDNA (Complementary Deoxyribose Nucleic Acid) as And step 2, screening out candidate reference genes of the purple perilla by using the sequencing data of the early-stage purple perilla transcriptome, namely PfActin, Pf18S, PfEF-1alpha, PfCYP, PfTUA, PfTUB and PfGAPDH respectively, and screening out the candidate reference genes of the purple perilla, namely PfActin, Pf18S, PfEF-1alpha, PfCYP, PfTUA, PfTUB and PfGAPDH. According to the method, the most stable reference genes in perilla leaves, stems, roots, fruits in three development stages and perilla leaves treated by 20% PEG6000 for 0H, 3H, 6H, 9H, 12H and 24H are screened from seven candidate reference genes, and a foundation is laid for deep excavation of functional genes in the later period.

The invention belongs to the technical field of plant biology, and particularly relates to a gene for regulating and controlling powdery mildew resistance of cucumbers and application of the gene, two cswrky31 mutants are obtained based on a cucumber Tnt1 reverse transcription transposon mutant library, and the two cswrky31 mutants both show high susceptibility to powdery mildew bacteria. A target gene Csa5G551250 which is directly regulated and controlled by CsWRKY31 is screened by virtue of a high-throughput transcriptome sequencing technology and a DNA affinity purification sequencing technology in combination with a yeast one-hybrid experiment. A dual luciferase report experiment shows that the CsWRKY31 positively regulates and controls the expression of the Csa5G551250, and the overexpression of the Csa5G551250 gene promotes the resistance of the cucumber to powdery mildew bacteria. A new reference gene resource is provided for cultivation of cucumber disease-resistant varieties, and a new thought is provided for molecular mechanism research of transcription factor mediated plant immunity.

The technology disclosed relates to detecting gene conservation and expression preservation. In particular, the technology disclosed relates to detecting gene conservation and epigenetic signals for a reference genetic sequence in comparison to a variant of the reference genetic sequence at base resolution through the generation of a plurality of alternative representations of the sequence in chromatin form which may represent evolutionary conservation, transcription initiation, or epigenetic signals, mapping the plurality of alternative chromatin sequences to a gene expression alterability classifier to generate a gene expression class prediction for the variant, and mapping the alternative chromatin sequence to a pathogenicity predictor to detect pathogenicity of variants.

The invention belongs to the field of pepper breeding, and discloses a KASP molecular marker closely linked with pepper fruit length, a primer, a kit and application, the molecular marker takes a Ca591.0 version gene as a reference gene, the single nucleotide polymorphism at the 70238018 position of the pepper No.3 chromosome is subjected to base A-T replacement, the flanking nucleotide sequence is shown as SEQ ID No: 1, the nucleotide sequence is shown as SEQ ID No: 2, and the nucleotide sequence is shown as SEQ ID No: 3. The sequences of the primers are shown as SEQ ID No: 2-4, and the kit comprises the primers. The invention also discloses an application of the primer or the kit in identification of different length types of pepper fruits or in molecular assisted breeding of pepper fruit length. A primer and a kit are developed aiming at the molecular marker, and can be used for identifying the length of the pepper fruit, so that assistant breeding is carried out by depending on the molecular marker, and the problems that the conventional breeding period is long and is easily influenced by the environment can be effectively solved.

The invention discloses a method for evaluating the detection effect of a fusion gene detection method, and belongs to the technical field of evaluation based on the detection effect. According to the invention, a detection reference substance I and a detection reference substance II are constructed, and the detection reference substances are connected with a reference gene segment in equal molecular number proportion, so that the detection condition under the condition of equivalent reference gene expression level can be realized, and whether the sample quality reaches the detection standard or not can be deduced; the measurement of the lowest detection quantity of the fusion gene relative to the expression quantity of the reference gene is realized; the first detection reference is used for detecting sensitivity analysis of the fusion type by the detection method of the fusion gene to be detected, and the second detection reference is used for verifying whether a detection result of the detection method of the fusion gene to be detected is a false negative result or not. And the method has a very good application prospect.

The invention belongs to the field of biotechnology application, and particularly relates to a goose whole genome liquid phase chip and application thereof. The SNP sites on the liquid phase chip are located in a goose reference genome GCA013030995.1, and the number of the SNP sites is 30000. The chip can specifically identify the genetic relationship between commercial geese and local geese, and can also be applied to whole genome association analysis, whole genome selective breeding, target trait QTL positioning analysis, population genetics analysis and the like. According to the invention, by capturing the reference gene and loci in the real sequence thereof, the coverage of sequencing data is improved, and the application value of the chip is enhanced.

The invention provides a probiotic screening method based on a fine tuning DNABERT model and a convolutional neural network. The method comprises the following steps: acquiring a reference genome sequence and a target sequence sample; preprocessing the reference genome sequence and the target sequence sample; segmenting the preprocessed reference gene sequence, and inputting the segmented reference gene sequence into a DNABERT model for fine tuning training; expanding the number of samples of the preprocessed target sequence, segmenting the sequence by using a sliding window, and inputting the segmented sequence into the fine-tuned DNABERT model for decoding to obtain a representation vector; inputting the representation vector into a CNN model for training, sequencing a target sequence to obtain a to-be-detected sequence sample, encoding the to-be-detected sequence sample, inputting the encoded to-be-detected sequence sample into the CNN model, judging whether the strain is the probiotics or not, and completing screening of the probiotics; according to the method, the data enhancement technology of introducing the reverse complementary sequence and the randomly intercepted sub-sequence is adopted, so that the diversity and the quantity of training data are increased, and the overfitting problem is effectively relieved.

The invention belongs to the technical field of biological detection, and discloses a primer probe composition for detecting CMV virus, a product and application thereof, the primer probe composition for detecting CMV virus comprises a primer probe combination for detecting CMV virus and a primer probe combination for detecting reference gene; the detection technology can be used for detecting the CMV on the DNA level and also can be used for detecting on the RNA level, the detection operation is more flexible, and the detection technology is particularly suitable for rapid safety recheck of cell therapy products and process detection in the production process. The primer probe composition provided by the invention has good detection sensitivity and specificity, is simple to operate, and can be widely applied to CMV detection of cell therapy products.

The invention relates to the technical field of molecular biology, in particular to a reference gene for Chinese olive functional gene expression analysis and application thereof. According to the invention, the reference gene RPN2B which is most stably expressed in fruits of different varieties (lines) of Chinese olives in different development and maturation periods is screened out, and in addition, two reference genes NIFS1 and RPS16 which are only second to the RPN2B are also screened out, so that the method can be used for expression analysis of nutritional quality or other functional genes of the fruits of the Chinese olives of different varieties (lines) in different development and maturation periods; the specificity, the stability and the accuracy of functional gene expression analysis data are remarkably improved, and powerful support is provided for Chinese olive functional gene expression analysis. The invention also provides a special detection primer for detecting the reference gene of the Chinese olive fruit, and the detection primer can quantify the expression quantity and function analysis of the auxiliary gene through fluorescence in the Chinese olive function research, and has high application value.

The application provides a urine exosome-based bladder cancer gene detection system and application, and relates to the technical field of biological detection. Through the cooperative application of the internal reference gene and the diagnosis model, the application provides a reliable endogenous quality control standard for the bladder tissue-derived exosome in the urine exosome sample; meanwhile, the application realizes non-invasive, accurate, rapid screening, diagnosis, prognosis and recurrence monitoring of bladder cancer, lays a key technical foundation and detection tool for the urine exosome-based molecular diagnosis of bladder cancer, and has important clinical transformation value and application prospect.

Provided is an internal reference gene for the real-time quantitative PCR (RT-qPCR) of Yersinia enterocolitica, including one of or a combination of some of glnS, nuoB, glmS, gyrB, dnaK, and thrS genes. Further provided is a primer set for amplifying the internal reference gene for the RT-qPCR of Yersinia enterocolitica. Given that stable housekeeping genes of Yersinia enterocolitica remain unclear, internal reference genes with relatively stable expression at different culture temperatures are selected by screening for Yersinia enterocolitica. Specific detection primers and real and reliable data provide a reference basis for selecting housekeeping genes of Yersinia enterocolitica; moreover, the use thereof alone or in combination, as an internal reference gene, improves the data accuracy, stability, and reliability, thereby solving the problem that usually only empirically selecting 16s rRNA as a single internal reference gene in conventional RT-qPCR results in unstable research results.

The invention discloses a molecular marker combination for detecting adulterated bovine milk in buffalo milk and application of the molecular marker combination. The molecular marker combination comprises a molecular marker 1 and a molecular marker 2, the molecular marker 1 is located at the 2552nd base of a buffalo reference gene sequence NC049568 or the 2540th base of a bovine reference gene sequence AB074968, and the polymorphism of the molecular marker 1 is T or C; the molecular marker 2 is located at the 2555th basic group of a buffalo reference gene sequence NC049568 or the 2543th basic group of a bovine reference gene sequence AB074968, and the polymorphism is G or A. The molecular marker 1 is a molecular marker 1. The molecular marker combination provided by the invention can be effectively used for detecting adulterated bovine milk in buffalo milk.

The invention belongs to the technical field of biological medicines, and particularly discloses a primer probe combination and a kit for detecting genotypes of SMA carriers based on a chromosome level and application of the primer probe combination and the kit. Wherein the primer probe combination comprises a primer probe group for detecting an exon 7 of an SMN1 gene, a probe for detecting an exon 7 of an SMN2 gene, a primer probe group for detecting a short-arm reference gene of a chromosome 5, a primer probe group for detecting a long-arm reference gene of the chromosome 5 and a primer probe group for detecting a reference gene of a human genome. The primer probe combination can eliminate chromosome copy number variation interference, greatly improves the specificity of SMA carrier genotype detection, can simultaneously detect the type I and type II of the SMA carrier genotype, has the detection sensitivity of 99%, and is suitable for large-scale population screening.

The present invention discloses lncRNA internal reference genes for osmanthus hormone treatment, primers, and applications thereof. The present invention obtains the optimal lncRNA internal reference genes for osmanthus under hormone treatment conditions: lnc00239991+18S under ABA treatment, lnc00265419+lnc00249739 under MeJA treatment, lnc00229717+lnc00044331 under ethephon treatment, and lnc00265419+lnc00239991 under multiple hormone treatments. qRT-PCR primers were designed using these genes. This invention fills the technical gap of the lack of internal reference lncRNAs under osmanthus hormone treatment, provides strong support for the quantification of lncRNAs under osmanthus hormone treatment, and provides a theoretical basis for the study of gene function in osmanthus.

The invention belongs to the technical field of forensic medicine, and particularly relates to an mRNA marker combination, a product and application thereof and a forensic human menstrual blood or human menstrual blood spot detection method. The mRNA marker combination comprises a reference gene, a human blood specific mRNA marker and a human menstrual blood specific mRNA marker; the human blood specific mRNA marker is composed of HBA, HBB, ALAS2 and GYPA; the human menstrual blood specific mRNA marker is composed of MMP7, MMP10, SFRP4 and STC1. Standardized identification suggestions are given according to mRNA marker combination expression conditions, the problems of insufficient specificity, flow disorder and the like in the prior art are solved, human menstrual blood or human menstrual blood spots can be accurately distinguished from peripheral blood and other body fluids, cross-platform compatible standardized detection is realized, evidence support is provided for missing cases, and the detection method is suitable for clinical application. The method has important practical significance for improving the forensic physical evidence identification efficiency.

According to the DUX4-IGH rearrangement detection primer probe set, the kit and the application, common rearrangement fracture sites of DUX4-IGH are confirmed, an upstream primer is designed on the upstream of a DUX4 transcript, 33 downstream primers are designed according to IGH fracture positions, a universal primer is designed on the upstream of DUX4, and a universal DUX4 probe is designed near the downstream of the primer; interference between the primer probes is small, influence of non-specific amplification is small, the primer probes are suitable for specific detection of multiple DUX4-IGH rearrangement sites, the problem that rearrangement detection is difficult in the prior art is solved, DUX4-IGH fusion transcripts and ABL1 reference genes can be quantitatively detected in a digital PCR instrument through RT-PCR, a basis is provided for auxiliary diagnosis, and the primer probes have remarkable application prospects.

The invention discloses a detection kit for detecting JAK2V617F mutation, which is characterized by comprising: a JAK2V617F detection reagent, which comprises an inner primer pair, an outer primer pair and a specific probe for specifically amplifying a JAK2V617F mutant gene; the internal reference detection reagent comprises a primer pair and a probe of an internal reference gene; the PCR reaction buffer solution comprises a reaction mixed solution and a HotstartTaq enzyme; the kit comprises a positive control, a known proportion of V617F mutant DNA, a negative control, a JAK2V617F wild type DNA, a positive control and a known proportion of V617F mutant DNA. Compared with the prior art, the invention has the advantage of providing the detection kit for detecting JAK2V617F mutation, which can be used for diagnosis, identification and clinical prognosis of PV.

The invention relates to the technical field of helicobacter pylori multi-drug-resistant gene detection, and discloses a paper sensor-based helicobacter pylori multi-drug-resistant gene detection and treatment quantitative monitoring kit, which comprises an isothermal amplification module, which comprises a loop-mediated isothermal amplification reaction system and is used for carrying out isothermal amplification on 23S rRNA gene and rdxA gene of helicobacter pylori; the paper sensor module comprises a nitrocellulose membrane, the nitrocellulose membrane comprises a test area and a control area, a biotin labeled probe aiming at 23S rRNA gene and rdxA gene mutant amplification products is fixed in the test area, and a biotin labeled probe aiming at reference gene cgt amplification products is fixed in the control area; and the color development module is used for performing color development and quantitative analysis on the detection result of the paper sensor module. The kit disclosed by the invention can be used for detecting various helicobacter pylori multi-drug-resistant genes, can be used for quantitatively monitoring, and is rapid and convenient to monitor.

The present invention relates to methods for determination of vector copy number. In particular, there is provided a method of determining a vector copy number in a biological sample that has undergone transduction with two or more vectors, the method comprising: (a) extracting genomic DNA (gDNA) from the biological sample; (b) performing digital droplet PCR (ddPCR) on the extracted gDNA using one or more pairs of forward and reverse primers to amplify a transgene of interest on each vector and to amplify a reference gene, together with one or more probes for detection of the amplification products; and (c) determining vector copy number for each vector in the biological sample.

The application belongs to the field of plant biotechnology, and particularly relates to a cucumber CsODO1 gene and application thereof, and is a MYB transcription factor capable of being combined with a CsCSE1 promoter, which is screened through a yeast single hybridization. It is proved through EMSA and tobacco transient expression analysis that the CsODO1 gene directly and specifically combines with a "CAACCA" sequence in the CsCSE1 promoter, and inhibits the expression of the CsCSE1 gene. The CsODO1 gene can be induced to express by a powdery mildew fungus, and is differentially expressed in the cucumber infected by the powdery mildew fungus. It is found through a cucumber cotyledon transient transformation system mediated by an agrobacterium that the CsODO1 gene negatively regulates the biosynthesis of lignin and the resistance of the cucumber to the powdery mildew fungus. The application provides a new reference gene resource for the cultivation of a cucumber disease-resistant variety, and provides a new thought for the research on the molecular mechanism of the plant immunity mediated by the lignin pathway.

The present invention relates to a c-myc gene copy number microdroplet digital PCR detection kit, belonging to the field of biomedicine technology. The present invention provides a c-myc gene copy number microdroplet digital PCR quantitative detection method, which calculates the c-myc gene copy number based on the fluorescence values of the c-myc gene and an internal reference gene; the present invention discloses primers and probes for the above method and kit. By combining the present invention with clinical pathological information, follow-up results, treatment response, and fluorescence in situ hybridization methods, the optimal threshold value of c-myc gene copy number amplification variation is determined, which serves as a reference indicator for the prognosis of specific tumor patients, providing a convenient and reliable auxiliary diagnostic tool for clinical practice.

The invention discloses a subsequence searching method, a comparison method, a system and equipment of a gene sequence. The subsequence searching method comprises the following steps: acquiring a preset number of sequencing data of a to-be-searched gene sequence; sequentially acquiring basic group data of a search interval corresponding to each piece of sequencing data on the reference gene sequence, and searching based on an acquisition result; the search interval corresponding to each piece of sequencing data is determined based on the to-be-searched base sequence of each piece of sequencing data; repeatedly obtaining the base data of the search interval of the next search, and searching based on the obtained result to obtain a target sub-sequence; the search interval of the next search is determined according to the next to-be-searched alkali of each piece of sequencing data and the search interval of the previous search. According to the search method, the basic group data of the search interval is obtained in advance, the sequencing data is processed in batches, calculation of other sequencing data can be processed when the prefetch result is waited, and the gene search efficiency is improved.

The invention belongs to the technical field of molecular biology, and particularly relates to a method for detecting transgenic source soybeans in soybean oil through an LAMP-OSD probe. The invention provides a rapid, sensitive, specific and visual method which is used for detecting whether soybean oil contains transgenic soybean components or not and a small amount of fragmented DNA (deoxyribonucleic acid) in the soybean oil, and detecting the transgenic soybean components in the soybean oil by simultaneously or respectively detecting two most common transgenic selection marker elements, namely a CaMV 35S promoter and an NOS terminator and a soybean reference gene Lectin. And whether the transgenic soybean is used in the raw material of the soybean oil can be effectively identified.

The application discloses a KASP molecular marker closely linked with pepper stem tip color, takes a zhangshugang version gene as a reference gene, and the KASP molecular marker is at 148515563 of a pepper 10th chromosome, a single nucleotide polymorphism at the position, and a base C to A replacement occurs at the position. The application further discloses primers, a kit for identifying the KASP molecular marker closely linked with the pepper stem tip color, and application of the KASP molecular marker in identifying the pepper stem tip color or identifying purity of pepper seeds. The KASP molecular marker closely linked with the pepper stem tip color gene, and the primers and the kit developed according to the KASP molecular marker can be used for identifying the pepper stem tip color and identifying the seed purity, can be detected at a seedling stage, and the identification method is carried out indoors, so that influences of natural factors such as weather, diseases and insect pests are avoided, and true and false hybrid seeds can be identified early, fast and accurately, and the seeds can be put on the market early.

The present invention discloses a genome for detecting the activation of multiple interferon pathways and its corresponding primer set, including primers for amplifying the genes of IFI44, MX1, IRF1, and GBP1. By combining the detection primer set with the internal reference gene HPRT1, a multiplex detection method has been developed, which can simultaneously perform accurate quantitative analysis on genes in patient samples, thereby achieving the typing of type I / II interferon pathways in one reaction and having high specificity and sensitivity. The detection method developed by the present invention simultaneously detects four target genes. Through clinical verification, its quantitative results are highly consistent with the results of separately detecting the four genes, greatly improving the detection efficiency while ensuring the reliability of the quantitative results. Combining the specificity of the screened genes and the reliability of the quadruple quantitative detection, the present invention can accurately identify the molecular mechanism causing autoimmune diseases and evaluate the development of the disease, providing effective guidance for clinical diagnosis and precision treatment.

The invention belongs to the field of molecular biology of forestry insects, and particularly relates to a fluorescent quantitative reference gene of different gender adults of yellow shield elephant, as well as a primer and application of the fluorescent quantitative reference gene. According to the method, 8 candidate genes commonly used as reference genes are referenced and selected according to transcriptome sequencing data of the hedychium aureum, gene sequences of the 8 reference genes are identified, the stability of the candidate reference genes is evaluated through five algorithms, and the fluorescent quantitative reference genes suitable for the hedychium aureum adults with different genders are obtained. And a real-time fluorescent quantitative PCR (Polymerase Chain Reaction) primer of the candidate reference gene is designed. The detection primer provided by the invention is strong in specificity and high in amplification efficiency, fills up the current situation that no reference gene exists under adults with different genders of the hedychium aureum, provides a reliable analysis basis for expression quantification of related functional genes under the adults with different genders of the hedychium aureum, and has a good application prospect. A necessary earlier-stage foundation is laid for analysis of subsequent molecular mechanisms and mining of key functional genes, and repeatability, stability and reliability of functional gene research can be improved at the same time.

The invention discloses a reference gene combination stably expressed by hermetia illucens under different conditions and application thereof, reference genes stably expressed by hermetia illucens in different development stages, different tissues, different genders and different genotypes are obtained through screening, and the reference gene combination is suitable for correction of hermetia illucens RT-qPCR detection results under different experimental conditions. Meanwhile, the invention further provides amplification primers for detecting the hermetia illucens reference genes, and the amplification primers are high in specificity and amplification efficiency. According to the invention, an early-stage foundation is laid for the quantification of related functional genes of hermetia illucens under different experimental conditions and the related research of molecular biology.

The invention provides a primer design, a disease typing diagnosis method and device, a composition, a kit and application thereof, and the primer design method comprises the following steps: obtaining a variety of reference data including a TRDV reference gene sequence and a TRDJ reference gene sequence; aiming at each kind of reference data, performing the following operations: aligning a plurality of reference gene sequences according to sites, determining a conservative score list of each site, obtaining conservative scores of a plurality of conservative intervals according to the conservative score list of each site, selecting K conservative intervals with higher conservative scores, K being a natural number greater than or equal to 1, and selecting K being a natural number greater than or equal to 1; generating K groups of primer combinations aiming at the K conservative intervals; and screening the primers in the K groups of primer combinations, evaluating the screened K groups of primer combinations, and obtaining a final primer combination according to an evaluation result.

The invention discloses screening and application of a fluorescent quantitative reference gene in a root and stem development process of iris vetiveria, and belongs to the technical field of plant genetic engineering. By constructing a strict reference gene evaluation and screening system, the reference gene with the most stable expression in different stages of root and stem development of iris vetiveria is screened out from eight candidate genes for the first time, errors in samples and between samples caused by internal reference selection are remarkably reduced, accurate correction and standardization of the expression level of a target gene are achieved, and the accuracy of the reference gene is improved. And reliable technical guarantee is provided for subsequent large-scale accurate quantitative analysis of irisone synthesis related functional genes.