87 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 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 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.

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 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 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 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 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.

The application discloses application and primers of CuSUMO1 and CuHTR4 as internal reference genes, belongs to the technical field of plant molecular biology, and particularly relates to application of CuSUMO1 and / or CuHTR4 as internal reference genes for quantitative analysis of genes of Dendrocalamopsis oldhami in Jinfoshan Mountain, and is characterized in that the nucleotide sequence of the CuSUMO1 is shown as SEQ ID NO. 2, and the nucleotide sequence of the CuHTR4 is shown as SEQ ID NO. 3. The application provides the internal reference genes CuSUMO1 and CuHTR4 and a primer pair for detecting expression analysis research of target genes of samples in different tissues, different development periods and different stress treatments of Dendrocalamopsis oldhami in Jinfoshan Mountain. The internal reference genes CuSUMO1 and CuHTR4 have high stability, and have important values for evaluating and detecting expression amounts of target genes of Dendrocalamopsis oldhami in Jinfoshan Mountain in different tissues, different development periods and different stress treatments.

The invention discloses a metagenome activity quantification method based on an exogenous artificially synthesized endogenous reference gene and application of the metagenome activity quantification method, and relates to the technical field of biology. According to the invention, an endogenous reference gene as shown in SEQ ID NO.1 is firstly designed, and the endogenous reference gene has no homology with an existing organism and can realize homology interference. On the basis, a metagenome activity quantitative method is further designed, and the core thought of the metagenome activity quantitative method comprises the following steps: providing an artificially synthesized vector containing an endogenous reference gene, setting the artificially synthesized vector as a standard system with gradient concentration, and adding the standard system into a sample to be detected; before addition, a to-be-detected sample is pretreated through light-sensitive DNA combined with dye so as to shield dead bacteria DNA interference, then nucleic acid extraction, library construction and sequencing are carried out, and metagenome activity quantification is obtained through bioinformatics analysis and calculation. According to the method, homology interference is eliminated from the source, absolute quantification of microbial activity is realized, the stability and standardization degree of an internal standard are improved, and the method is suitable for quantitative analysis of metagenomes in various scenes.

The invention discloses a method for synchronously detecting multiple strawberry pathogens and application, the method for synchronously detecting multiple strawberry pathogens comprises a primer combination capable of synchronously detecting multiple strawberry pathogens, and the primer combination comprises 18 specific primers and 2 universal primers, the fluorescent probe can be used for detecting strawberry mottle virus, strawberry mild yellow edge virus, strawberry crinkling virus, strawberry vein banding virus, strawberry powdery mildew, strawberry gray mold, strawberry anthracnose, strawberry xanthomonas and strawberry internal reference beta-Actin1, and also provides a corresponding detection method, a detection reagent and a detection kit. According to the method, 9 targets can be detected at the same time through one-tube reaction, the detection efficiency is greatly improved, meanwhile, a false negative result is effectively avoided through internal reference gene monitoring, powerful technical support is provided for early diagnosis of strawberry diseases, and the method has great application value and market prospects.

The invention provides a primer design, a disease typing diagnosis method and device, a composition, a kit and application thereof. The primer design method comprises the following steps: acquiring various reference data, wherein the various reference data comprise a TRGV reference gene sequence and a TRGJ 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.

Provided are a method and device for primer design, a method and device for disease typing diagnosis, a composition, a kit and the use thereof. The method for primer design comprises: acquiring one or more pieces of reference data, each piece of reference data comprising a plurality of reference gene sequences; and for each piece of reference data, performing the following operations: aligning the plurality of reference gene sequences by sites, determining a conservation score list for each site, obtaining conservation scores of a plurality of conservation intervals on the basis of the conservation score list of each site, selecting K conservation intervals with relatively high conservation scores, K being a natural number greater than or equal to 1, and generating K sets of primer combinations for the K conservation intervals; and screening primers in the K sets of primer combinations, evaluating the screened K sets of primer combinations, and obtaining a final primer combination on the basis of an evaluation result.

Provided are a primer design method and device, a disease subtyping diagnosis method and device, a composition, a kit, and a use thereof. The primer design method comprises: acquiring multiple types of reference data, the multiple types of reference data each comprising a TRDV reference gene sequence and a TRDJ reference gene sequence; and performing the following operations on each type of reference data: aligning a plurality of reference gene sequences site-by-site to determine a conservation score list for each site, obtaining conservation scores for a plurality of conservation intervals on the basis of the conservation score list for each site, selecting K conservation intervals having relatively high conservation scores, wherein K is a natural number greater than or equal to 1, and generating K primer combinations for the K conservation intervals; and screening primers in the K primer combinations, evaluating the K primer combinations having undergone screening, and obtaining a final primer combination on the basis of the evaluation result.

The embodiment of the invention discloses a method for screening fluorescent quantitative reference genes of different parts of zingiber mahogany, relates to the technical field of gene detection, and aims to provide effective reference genes for future research on ornamental characters and physiological and biochemical gene functions of zingiber mahogany. The method for screening the fluorescent quantitative reference genes at different parts of zingiber mahogany comprises the following steps: extracting RNA (Ribonucleic Acid) of five parts, namely roots, leaves, lateral buds, bracts and florets of zingiber mahogany 'Bailong' to obtain candidate reference genes; carrying out qRT-PCR (quantitative reverse transcription-polymerase chain reaction) treatment on the candidate reference genes, comprehensively evaluating the expression stability of the candidate reference genes through BestKeeper, NormFinder and geNorm software programs, and screening out target reference genes with stable expression at different parts of the zingiber mairei 'Bailong'; the candidate reference genes comprise at least one of MDH (malondialdehyde), TAP46 (triphosphate synthase 46), EF1 alpha 2, Actin12, 18SrRNA (ribose nucleic acid), GAPDH (glyceraldehyde-phosphate dehydrogenase) and TUB7 (triphosphate synthase 7).

The application provides a high-collagen trait-associated SNP marker combination of Pseudosciaena crocea and breeding application. Specifically provided is application of a chip loaded with the high-collagen trait-associated SNP marker combination of Pseudosciaena crocea in high-collagen trait genotyping or high-collagen trait breeding, and the SNP marker combination is composed of 12 SNP molecular markers, the location of each SNP molecular marker on a chromosome, an altered allele, and reference gene information are shown in Table 1. The SNP marker combination of the application can be used to determine the genetic potential of collagen content of individuals in early offspring, and the best individuals with high collagen potential of a certain scale can be selected in advance, and the selected individuals are bred and cultivated to parents, only the target individuals are bred and cultivated subsequently, and a large number of offspring seedlings do not need to be bred and cultivated in the breeding process, so that the breeding cost can be greatly reduced.

The invention discloses a sheep reference gene MTPN and application thereof. According to the application of the sheep reference gene MTPN in RT-qPCR detection of the expression level of the target gene of the sheep early embryo, the reference gene MTPN can be independently used, and the sheep reference gene MTPN and the sheep reference gene Ppia can also form an RT-qPCR internal reference combination to detect the expression level of the target gene of the sheep early embryo. According to the invention, proper reference genes are screened for gene expression research of sheep early embryos at different development stages, and a powerful guarantee is provided for the accuracy of real-time fluorescent quantitative PCR detection of the gene expression level of the sheep early embryos.

The invention belongs to the technical field of plant biology, and particularly provides application of a CsTSR gene in cucumber corynespora leaf spot resistance. A transient expression strain and a cstsr mutant are respectively obtained through transient transformation of the CsTSR gene and a Tnt1 reverse transcription transposon mutant library, and the CsTSR gene is found to positively regulate the resistance of the cucumber to the corynespora leaf spot disease. The miR164d can regulate and control the CsTSR transcription factor in a targeted manner. A dual luciferase report and a GUS (glucuronidase) staining experiment result show that the miR164d negatively regulates and controls the CsTSR, and the CsTSR-mediated cucumber disease resistance can be enhanced by silencing the miR164d. The invention discloses a functional mechanism of the miR164d-CsTSR molecular module in cucumber pathogenic bacterium infection resistance, and provides a new reference gene resource for cultivation of cucumber disease-resistant varieties.

Provided are a method and device for primer design, a method and device for disease typing diagnosis, a composition, a kit and the use thereof. The method for primer design comprises: acquiring one or more pieces of reference data, each piece of reference data comprising a plurality of reference gene sequences; and for each piece of reference data, performing the following operations: aligning the plurality of reference gene sequences by sites, determining a conservation score list for each site, obtaining conservation scores of a plurality of conservation intervals on the basis of the conservation score list of each site, selecting K conservation intervals with relatively high conservation scores, K being a natural number greater than or equal to 1, and generating K sets of primer combinations for the K conservation intervals; and screening primers in the K sets of primer combinations, evaluating the screened K sets of primer combinations, and obtaining a final primer combination on the basis of an evaluation result.

The present application provides a method for quantitative detection of rice purity and a kit for quantitative detection of rice purity. The method for quantitative detection of rice purity provided by the present application comprises: 1) extracting rice sample DNA; 2) performing PCR amplification using primers that can amplify the sequence of the difference fragments of fragrant rice and non-fragrant rice; and 3) performing quantitative analysis to determine the purity of fragrant rice in the rice sample. The present application utilizes the specific sequence of fragrant rice that is different from non-fragrant rice, designs efficient and sensitive fragrant rice specific primers and probes, performs real-time fluorescent PCR amplification on sample DNA, and performs quantitative analysis on the amplification data by setting an endogenous reference gene and a reference sample, thereby quantitatively determining the purity of fragrant rice. The quantitative detection of rice purity based on molecular biology effectively solves the problem of quantitative identification of non-fragrant rice mixed in fragrant rice.

The application discloses a gene amplification detection method based on nucleic acid mass spectrometry technology, comprising the following steps: setting a target gene region, configuring a reference gene, and confirming experimental primers based on the target gene region and the reference gene; obtaining an experimental sample, performing a primer reaction processing operation based on the experimental primers and the experimental sample, and obtaining a sample to be analyzed; setting an interpretation interval, configuring a sample applicator and a nucleic acid mass spectrometer, performing a data analysis and interpretation operation based on the sample to be analyzed, the interpretation interval, the sample applicator and the nucleic acid mass spectrometer, and obtaining a gene amplification interpretation result; the application can achieve a detection accuracy higher than that of the fluorescence in situ hybridization technology and the immunohistochemical technology in the prior art, a detection throughput higher than that of the fluorescent quantitative PCR technology and the digital PCR technology, a detection cost lower than that of the fluorescent quantitative PCR technology and the digital PCR technology, and a detection time shorter than that of the second-generation sequencing NGS technology.

The present application relates to the internal reference gene of 'bean green' peony and its primer and application, and belongs to the field of biotechnology, the present application takes green peony 'bean green' as the test material, analyzes the expression stability of 9 candidate internal reference genes, and the results show that UBC and MBF1A In different tissues of 'bean green'; GAPDH and ACT In different development periods of 'bean green' petal stamen, the present application constructs a stable and reliable quantitative detection system of 'bean green', provides a basis for gene expression analysis in 'bean green' peony and the molecular mechanism research of 'bean green' flower type and flower color formation, provides technical support for prolonging the coloring time of 'bean green' flower and improving its ornamental quality, provides a reference for the green flower research of other varieties of green peony, and also provides a theoretical basis for the subsequent cultivation of new varieties of green flower peony.

The present application relates to a primer combination for lung cancer detection and its application in lung cancer diagnosis, in particular, a technology for specifically detecting CIZ1b splice body RNA through real-time fluorescent quantitative PCR technology. The method combines blocker sequence to inhibit non-specific amplification of CIZ1a splice body by designing specific primers and probes, and ensures high-specificity detection of CIZ1b splice body RNA. The detection samples include lung cancer tissues and plasma, and cDNA is generated through reverse transcription after RNA extraction of the samples, and real-time fluorescent quantitative analysis is performed. Meanwhile, the method introduces an internal reference gene B2M for standardization correction, and improves the sensitivity and accuracy of the detection. The present application provides a non-invasive and reliable early lung cancer diagnosis tool, which has good clinical application potential.

The invention discloses a peony flowering phase regulation PCR detection reference gene, a primer, a screening method and application, and belongs to the technical field of plant molecular biology, the reference gene is RPL17, and the nucleotide sequence is shown as Seq ID No. 25 in a sequence table. And specific primers for amplifying the reference gene RPL17 are shown as Seq ID No. 17 and Seq ID No. 18 in a sequence table. The gene can meet the requirements of qRT-PCR (quantitative reverse transcription-polymerase chain reaction) technology for detecting the expression level of related genes of white peony varieties in the peony development process and in different flowering periods, and the stability and the accuracy for detecting the expression of the related genes of the white peony varieties in the peony development process and in different flowering periods are improved; the difference between samples and the difference inside the samples caused by selection of the reference genes are reduced to the maximum extent, and a solid foundation is laid for later analysis of expression modes of key genes related to peony flowering phase regulation by using qRT-PCR (quantitative reverse transcription-polymerase chain reaction).