1887 results about "Genomic clone" patented technology

Provided herein are methods, compositions, and kits to spatially detect a polypeptide-nucleic acid complex or a protein-nucleic acid complex in a biological sample. For example, such methods can include contacting binding agents with a biological sample, wherein a binding agent specifically binds a polypeptide of a polypeptide-nucleic acid complex; aligning the biological sample with a substrate comprising a plurality of capture moieties, wherein the binding agent interacts with a capture moiety; releasing the nucleic acid of the binding agent-polypeptide-nucleic acid complex; hybridizing the released nucleic acid to a capture domain of a capture probe on an array; and using determined sequences of a spatial barcode in the capture probe and the released nucleic acid to determine the location of the released nucleic acid in the biological sample, thereby determining the location of the polypeptide-nucleic acid complex in the biological sample.

Disclosed herein are methods of amplifying an analyte in a biological sample using a bridging oligonucleotide that hybridizes to a captured analyte. The methods disclosed herein include steps of (a) contacting a biological sample with a substrate having capture probes comprising a capture domain and a spatial barcode; (b) hybridizing the analyte to the capture domain; and (c) contacting the analyte to a bridging oligonucleotide comprising (i) a capture-probe-binding sequence, and (ii) an analyte-binding sequence; (d) extending the bridging oligonucleotide; and (e) determining (i) all or a part of the sequence of the analyte, or a complement thereof, and (ii) the spatial barcode, or a complement thereof, and using the determined sequence of (i) and (ii) to determine the location of the analyte in the biological sample.

The invention discloses a sugarcane germplasm resource evaluation and breeding method for smart agriculture. The sugarcane germplasm resource evaluation and breeding method comprises the following steps: step 1, collecting materials with wide genetic diversity; establishing an intelligent incubator and a greenhouse, and monitoring environmental parameters by using an Internet of Things sensor; 2, performing data acquisition on the related germplasm resources by using an unmanned aerial vehicle, a robot and a near infrared spectrum technology, and constructing a phenotype database for data management; meanwhile, genotype identification is carried out by combining molecular marker-assisted selection and genome selection technologies, molecular markers of key genes are mined, and the genetic value of related germplasm is evaluated; step 3, constructing a hybrid combination prediction model by using an AI algorithm, optimizing parent matching of the related germplasm and the cultivated sugarcane, monitoring and collecting the filial generation at the same time, and analyzing and screening in combination with smart agriculture; 4, demonstration planting is conducted on the bred excellent sugarcane strain, and application of the excellent strain is improved through government-enterprise-scientific research institution collaborative popularization and in combination with farmer technical training.

The invention relates to the technical field of crop genetic breeding and plant molecular design breeding, in particular to a pakchoi whole genome liquid phase chip and application. The pakchoi breeding gene chip disclosed by the invention comprises a target capture probe which is designed by taking 1000 core SNP (Single Nucleotide Polymorphism) molecular markers positioned on 40KSNP sites on a pakchoi reference genome version V4.0 as templates. The pakchoi whole genome liquid phase chip provided by the invention comprises a 40K site targeted capture probe mixed solution and a hybridization capture reagent. The pakchoi 40KSNP targeted capture probe can rapidly and effectively track genetic materials of pakchoi, and is suitable for different application scenes such as accurate identification of pakchoi germplasm resources, genetic relationship analysis between materials, variety authenticity identification, QTL gene positioning and mining, molecular marker-assisted breeding, variety improvement and whole genome selective breeding application. Therefore, the method has good application prospect and important social value.

The invention discloses a tea tree liquid phase chip and application thereof, and relates to the technical field of molecular detection. The invention discloses a tea tree liquid phase chip and application thereof, according to site screening requirements and probe design principles, the tea tree liquid phase chip comprises 5781 SNP sites, and tea tree resource genetic typing can be realized based on a target interval genome sequence liquid phase capture accurate positioning sequencing typing technology. The tea tree liquid chip can realize low-cost genetic typing, is mainly specific to tea trees, can realize variety identification and genetic relationship analysis of the tea trees, scientifically guides hybridization improvement work of the tea trees and assists protection and development of germplasm resources of the tea trees, and has relatively high application values in multiple fields of tea tree breeding. The tea tree liquid phase chip can be used for tea tree genetic diversity evaluation, germplasm resource and genetic relationship identification, genetic map construction and gene localization, whole genome association analysis and tea tree molecular marker assisted breeding.

The present invention, among other things, provides technologies for detecting and / or quantifying nucleic acids in cells, tissues, organs, or organisms. Pre-designed barcodes are associated with specific molecular targets through sequential hybridization experiments. A pseudo-color based barcoding scheme is described that overcomes the limitations in the previous generation of the technology. The current method can be applied to both in vitro and in situ analysis.

The invention belongs to the technical field of molecular biology, and particularly relates to a CeNF-YA3 gene, an expression vector and application of the CeNF-YA3 gene in vegetable fat regulation and control. The nucleotide sequence of the CeNF-YA3 gene is as shown in SEQ ID NO.1. After the CeNF-YA3 gene is over-expressed, the grease content of arabidopsis seeds and leaves can be remarkably increased; in addition, the invention also provides a series of vectors containing the CeNF-YA3 gene for subcellular localization, yeast hybridization and plant overexpression for the first time, and proves that the CeNF-YA3 protein has a transcriptional activation function, is localized in a cell nucleus and accords with the basic characteristics of transcription factors for the first time. Therefore, the technical scheme of the invention provides scientific guidance for the application of the CeNF-YA3 gene and the protein thereof.

The present disclosure provides compositions and methods that employ the compositions for conducting pairwise sequencing and for generating concatemer template molecules for pairwise sequencing. The concatemers can be generated using a rolling circle amplification reaction which is conducted either on-support, or conducted in-solution and then distributed onto a support. The rolling circle amplification reaction generates concatemers containing tandem copies of a sequence of interest and at least one universal adaptor sequence. An increase in the number of tandem copies in a given concatemer increases the number of sites along the concatemer for hybridizing to multiple sequencing primers which serve as multiple initiation sites for polymerase-catalyzed sequencing reactions. When the sequencing reaction employs detectably labeled nucleotides and / or detectably labeled multivalent molecules (e.g., having nucleotide units), the signals emitted by the nucleotides or nucleotide units that participate in the parallel sequencing reactions along the concatemer yields an increased signal intensity for each concatemer.

The invention discloses application of an upstream non-coding region uORF of an ABA2 gene in regulation and control of crop agronomic traits, in particular to application in improvement of pre-harvest sprouting resistance of rice and cultivation of pre-harvest sprouting resistant rice strains, and belongs to the technical field of gene engineering. The invention finds that uORF in the upstream non-coding region of the crop ABA2 gene can inhibit the translation level of downstream ABA2 protein on the basis of not influencing the transcriptional level, so that the uORF can participate in regulation and control of crop agronomic traits in which ABA participates in regulation and control, such as improvement of pre-harvest sprouting resistance of rice. Therefore, the upstream non-coding region uORF of the crop ABA2 gene can be modified through a hybridization or gene editing technology, so that the purpose of regulating and controlling crop agronomic traits in which ABA participates in regulation and control is achieved, for example, site-directed mutagenesis is performed on an initiation codon of the upstream non-coding region uORF of the rice ABA2 gene, and the pre-harvest sprouting resistance of rice is improved.

The invention discloses an artificial breeding method of epinephelus fuscoguttatus and pink pterus coerulescens hybrid species, which comprises the following steps: screening healthy epinephelus fuscoguttatus as a female parent, screening healthy pink pterus coerulescens as a male parent, and carrying out enhanced breeding on the male parent and the female parent, the gonad maturation of the parent is promoted; the method comprises the following steps: collecting egg liquid of a female parent and semen of a male parent, inseminating to obtain a fertilized egg, incubating the fertilized egg to obtain a hybrid spot, performing authenticity identification on the hybrid spot by using one or more of sequences SEQ ID NO.1-SEQ ID NO.8, and determining that the hybrid spot inherits genetic information of the female parent and the male parent. The technical blank of crossbreeding of the pink pink bass as the parent grouper is filled, so that the hybrid obtained by hybridizing the epinephelus fuscoguttatus and the pink pink bass is reliable in authenticity, and inherits genetic information of male and female parents on the molecular level.

Disclosed are a genomic mating method for Huaxi cattle based on whole genome single nucleotide polymorphism (SNP) information and an application thereof. The method includes the following specific steps: step 1, extracting deoxyribonucleic acid (DNA) from to-be-hybridized Huaxi cattle individuals for genotyping; step 2, performing genotype data imputation to obtain high-density chip data; step 3, calculating an additive genetic relationship matrix, utilizing genomic best linear unbiased prediction (GBLUP) to obtain genomic estimated breeding values of five important economic traits of a to-be-hybridized Huaxi cattle population, and calculating a comprehensive selection index of the individuals; and step 4, using a genetic algorithm to construct a population optimal mating combination list. In the present invention, the breeding cost is greatly saved and an inbreeding level of offspring populations is reduced.

The invention provides a method for constructing a rice epigenetic recombinant inbred line, and belongs to the technical field of molecular biology and genetic breeding. According to the method for constructing the rice epigenetic recombination inbred line, a fertile rice DDM1 gene function weakening mutant is obtained through a CRISPR / Cas9 gene editing technology and hybridized with wild type rice with the same background to obtain F1, then the F1 and the wild type rice are subjected to backcrossing to obtain BC1F1, single-plant offspring selfing with the DDM1 genotype being the wild type is selected, selfing is conducted for multiple generations through a single-grain transmission method, and the rice epigenetic recombination inbred line is obtained. According to the present invention, the strain population with stable inheritance is constructed, such that the rice epigenetic recombinant inbred line is obtained, and has characteristics of rich phenotype, genetic and epigenetic variation so as to provide important population resources for batch identification and the like of important agronomic shape functional sites of rice.

The invention belongs to the technical field of biosensors, and provides a cadmium telluride quantum dot-based fluorescent biosensor for detecting tetracycline, which comprises a CdTe quantum dot, an Apt-T composite probe, M-2P, a C chain and an Ag-HP chain, the Apt-T composite probe is obtained by hybridizing Apt and a T chain; the M-2P is obtained by hybridizing an M chain and a P chain according to a molar ratio of 1: 2; the nucleotide sequences of Apt, T, M, P, C and HP are as shown in SEQ ID NO: 1-6. The sensor disclosed by the invention has the advantages of high detection speed, simplicity in operation, low price, low detection limit, high specificity and the like, can make up for the defects and deficiencies of the existing tetracycline detection method, and realizes rapid and accurate quantitative detection of tetracycline.

The invention relates to the technical field of biology, in particular to a KASP marker for identifying corn haploids, a primer group and application of the KASP marker and the primer group. According to the invention, a corn Maize 60K chip is used for respectively hybridizing eight inbred lines (Mo17, B73, Qi319, HZ4, Z58, W64A, W22 and TX) with an induction line CAU5 to screen out eight groups of haploids (10 haploids are respectively induced in each group as repetitions) from filial generations F1, 145 sites which are shown as 100% stable completion of male parent chromosome elimination in respective 10 haploids of the eight F1 combinations are obtained, and for the 145 sites, the male parent chromosome elimination is completely completed. 94 stable KASP markers are developed, and a new way is provided for accurate screening of haploids.

The invention belongs to the technical field of molecular biology, and particularly relates to a CeNF-YA1 gene, an expression vector and application of the CeNF-YA1 gene in vegetable fat regulation and control. The invention provides the CeNF-YA1 gene for the first time, the nucleotide sequence of the CeNF-YA1 gene is as shown in SEQ ID NO.1, the full length of a coding region of the CeNF-YA1 gene is 792 bp, a series of carriers containing the CeNF-YA1 gene, such as subcellular localization, yeast hybridization and plant overexpression, are constructed for the first time, and meanwhile, the CeNF-YA1 protein is proved to have a transcription activation function, is localized in a cell nucleus and conforms to the basic characteristics of transcription factors. It is found for the first time that overexpression of the CeNF-YA1 gene can significantly improve the grease content of arabidopsis seeds and leaves; therefore, the technical scheme provided by the invention can be used for improving the oil content of the plant seeds and the nutritional tissues, and shows the application prospect in improving the oil content of the seeds and the nutritional tissues.

The invention provides a closely linked molecular marker primer of a rice brown planthopper resistant major gene Bph56 as well as a marking method and application of the closely linked molecular marker primer. According to the invention, genotypes of F2 individual plants obtained by hybridizing a rice insect-resistant variety C127 (or) and KW (or) are combined with brown planthopper resistance levels of an F3 family for genetic linkage analysis, a resistance major gene Bph56 carried by the insect-resistant variety C127 is identified, and a region of the resistance major gene Bph56 is reduced to a 77kb fragment limited by markers 6M26.278 and 6M26.355 on two sides and is closely linked with a molecular marker 6M26.355. The molecular marker 6M26.355 disclosed by the invention can be used for effectively detecting whether an insect-resistant variety C127 and derived varieties (lines) of the insect-resistant variety C127 contain the major resistance gene locus or not, so that the selection efficiency of the brown planthopper-resistant character is greatly improved, and the brown planthopper-resistant rice variety (line) containing Bph56 is obtained.

The invention discloses application of a gene CsPOD7 in cucumber haploid breeding, and belongs to the technical field of molecular breeding. According to the invention, a parthenogenesis haploid induction gene CsPOD7 is cloned from cucumber, the CsPOD7 is knocked out through a CRISPR-Cas9 gene editing technology, and a cspod7 mutant is used as a male parent to be hybridized with other cucumber materials as a female parent, so that parthenogenesis haploid can be generated. The invention is the first application of the gene CsPOD7 in developing a parthenogenesis haploid induction line in dicotyledon, realizes the application of the haploid induction line based on the gene CsPOD7 in cucumber genetic breeding, accelerates the cucumber breeding process, and has huge potential application value and market prospect.

The invention belongs to the technical field of microbiological detection, and discloses a composite probe for detecting salmonella enteritidis and a preparation method. The composite probe for detecting the salmonella enteritidis comprises magnetic nanoparticles FeO, wherein the surface of the magnetic nanoparticles FeO is modified with an aptamer apt which is specifically combined with the salmonella enteritidis; the surface of the long afterglow nano particle PLNPs is modified with a DNA (Deoxyribose Nucleic Acid) sequence cDNA (Complementary Deoxyribonucleic Acid) complementary with the aptamer; the aptamer apt and the complementary DNA sequence cDNA are subjected to hybridization to form a composite probe structure FeO-SEapt (at) PLNPs-cDNA (complementary deoxyribonucleic acid). The long afterglow luminescence characteristic of PLNPs (ZnGaO: Cr) is utilized, and an excitation light source is stopped before detection, so that signal acquisition completely avoids autofluorescence of a sample matrix, background fluorescence interference is thoroughly eliminated, and the signal-to-noise ratio is remarkably improved.

The present disclosure in some aspects relates to methods, compositions, and kits for rolling circle amplification (RCA) comprising extending a nucleic acid priming sequence hybridized to a circular nucleic acid template using a polymerase to generate an extended priming sequence, wherein the circular nucleic acid template is crosslinked to a second nucleic acid strand; de-crosslinking the circular nucleic acid template from the second nucleic acid strand; and extending the extended priming sequence to generate a rolling circle amplification product (RCP).

The invention relates to the technical field of biomedical engineering, and discloses a preparation method of a nano-enzyme SERS (Surface Enhanced Raman Scattering) sensor for detecting diabetic retinopathy, which comprises the following steps: (1) synthesizing a bimetallic gold-palladium nanorod; (2) assembling a gold trioctahedron array; and (3) modifying the surface of the gold-palladium nanorod synthesized in the step (1) with single-stranded DNA1 to prepare a nano-enzyme probe, hybridizing an aptamer of a vascular endothelial growth factor (VEGF) with single-stranded DNA2, modifying the surface of the gold trioctahedron array assembled in the step (2) with the hybridized aptamer, preparing a capture substrate, and constructing the nano-enzyme SERS sensor by the nano-enzyme probe and the capture substrate. In the detection process, the VEGF triggers a competitive recognition mechanism to promote the nano-enzyme probe to be captured by the capture substrate and catalyze tetramethyl benzidine (TMB) to generate oxTMB with a strong SERS signal. The sensor has the advantages of high sensitivity, strong specificity and good accuracy, and can realize rapid quantitative detection of DR biomarkers in serum.

The invention discloses a construction method and application of a mouse model for specifically tracing heart valve cells, and belongs to the technical field of animal model construction. The model construction method comprises the following steps: (1) constructing a Wid1-Cre hybrid mouse before inserting a Cre recombinase coding gene into a termination codon of a mouse Wid1 gene; and (2) hybridizing the Wid1-Cre hybrid mouse with the Rosa26-tdTomato homozygous report mouse, so as to obtain the double transgenic mouse which simultaneously carries the Wid1-Cre gene and the Rosa26-tdTomato gene. According to the invention, on the basis of the expression characteristic that Wif1 is only limited to a valve area in the heart, Wif1-Cre is constructed; according to the Rosa26-tdTomato mouse model, specific tracing of heart valve cells is achieved, and an important experimental tool is provided for mechanism research of heart valve diseases and development of therapeutic drugs.

Provided herein are methods, compositions, and kits for detecting analytes of interest from biological samples using tagmentation. The method comprises: (a) hybridising a nucleic acid analyte to a capture probe on an array, wherein the capture probe comprises: (i) a spatial barcode and (ii) a capture domain; (b) extending the capture probe using the nucleic acid analyte as a template, thereby generating a barcoded nucleic acid molecule on the array, wherein the barcoded nucleic acid molecule comprises a complement of the sequence of the nucleic acid analyte; and (c) tagmenting the barcoded nucleic acid molecule or a derivative thereof using a transposome comprising a transposase to insert a transposon end sequence into the barcoded nucleic acid molecule or the derivative thereof, thereby generating a barcoded nucleic acid fragment, wherein the barcoded nucleic acid fragment comprises: (i) the barcode, (ii) the transposon end sequence, and (iii) at least a portion of a sequence of the nucleic acid analyte or a complement thereof.

The invention discloses a male sterility gene ZmUCE4 and application thereof in creating a male sterility line of corn, and belongs to the field of plant biotechnology breeding. According to the invention, site-directed mutagenesis of the ZmUCE4 gene in wild type corn is carried out through a CRISPR / Cas9 gene editing technology, pollen development abnormality and abortion can be caused, complete male sterility is caused, and it is proved that the ZmUCE4 gene has a regulation and control function on corn male reproductive development. Through offspring screening, a sterile line without transgenic components can be obtained, a stable maize male sterile line is created, and the method has important significance on maize male fertility control and hybrid seed production. The invention also designs a functional molecular marker aiming at the obtained uce4 male sterility mutant gene, and has important application value in maize male sterility line cultivation, sterile hybrid seed production and molecular marker-assisted selection.

The invention relates to the technical field of life science, in particular to a method for constructing an atrophic gastritis-based SPEM lesion model, which comprises the following steps: S1, constructing an Slc7a11 conditional knockout mouse strain: carrying out cage hybridization on an Slc7a11 flox / flox mouse and an ATP4b-Cre transgenic mouse, and knocking out a gastric wall cell specific Slc7a11 gene; s2, carrying out genotyping identification; the identification method based on the atrophic gastritis SPEM lesion model and the construction method based on the atrophic gastritis SPEM lesion model comprise the following steps: S1, pathological model verification: taking a mouse gastric mucosa tissue slice, quantitatively analyzing the proportion of an SPEM region by adopting an immunofluorescence double-standard technology, and confirming Slc7a11 knockout induced SPEM pathological characteristics in combination with the existence of SPEM in the gastric mucosa; and S2, long-term pathology evolution monitoring. Through gastric wall cell specific Slc7a11 gene knockout and multiple PCR identification technologies, the initial response SPEM state after gastric mucosa damage is successfully simulated, and the kit is used for researching atrophic gastritis pathological mechanisms and evaluating drug intervention.

The invention discloses an SNP (Single Nucleotide Polymorphism) molecular marker related to corn plant height regulation and application thereof. The sequence of the molecular marker is shown as SEQ ID NO.1, the 201bp site basic group of the sequence from the 5th end shows C / T polymorphism, a temperate zone corn inbred line Ye107 with low plant height is used as a common male parent and is respectively hybridized with one temperate zone and four tropical / subtropical corn inbred lines as female parents, and a corn polyparent group with obvious plant height difference is constructed. GWAS analysis and linkage analysis are used for co-localization to Chr1: SNP-25, 296 and 496 which are located in a chromosome 1 and remarkably related to the plant height, 3.59% of plant height phenotypic variation can be explained through the loci, a functional gene for regulating and controlling the corn plant height is excavated from the loci, and a qRT-PCR result shows that the gene is highly expressed in corn V14 period internodes, so that it is proved that the gene participates in regulation and control height correlation of the corn plant height, and the functional gene is a functional gene for regulating and controlling the corn plant height. Results of the invention provide technical support for efficient and accurate breeding of high-stalk or dwarf high-yield corn varieties by molecular markers.

Methods for determining a location of a feature on an array include: (a) providing a first array with a first plurality of features immobilized on a first substrate; (b) providing a second array with a second plurality of features immobilized on a second substrate; (c) aligning the first array with the second array; (d) hybridizing a first barcoded oligonucleotide of the first array to a second barcoded oligonucleotide of the second array, thereby producing a combined nucleic acid that includes first and second spatial barcodes; (e) determining all or a portion of the sequence of the combined nucleic acid; and (f) identifying the second barcoded oligonucleotide associated with the first barcoded oligonucleotide in the combined nucleic acid, and determining the location of a second feature in the second array.

The invention relates to the field of hybrid rice breeding, and discloses a method for directionally breeding mutant hybrid rice parents. According to the method, mutation tillering is accurately positioned in the M1 tillering stage through the KASP technology, and a directional pollination / harvesting strategy is combined, so that the genetic bottleneck caused by chimeras and low fertility (especially sterile lines) is effectively overcome, the mutation transmission efficiency is maximized, and the M1 generation mutation transmission efficiency is remarkably improved. Through early accurate identification and efficient propagation of the M1 generation, homozygotes can be obtained and screened from the M2 generation, and compared with a traditional process, the method has the advantages that the obtaining process of low-cadmium homozygous parents is obviously accelerated, and the breeding period is greatly shortened. Heavy ion mutagenesis, targeted sequencing, high-throughput KASP typing and directional propagation technologies are integrated, and the whole process from mutagenesis to obtaining of the homozygous low-cadmium parent is rapid, accurate and efficient.

The invention provides a breeding method for constructing XX / XY sex determination all-female sterile fish and application. The method comprises the following steps: knocking out a catalytic enzyme coding gene of a fish sex steroid hormone synthesis pathway by using a gene editing technology to block the fish sex steroid hormone synthesis pathway, screening to obtain an effectively mutated homozygote XX sex genetic type pseudo male fish, and hybridizing the pseudo male fish with improved allotetraploid crucian and carp female fish to obtain the fish sex steroid hormone synthesis pathway. Therefore, an allotriploid population is obtained, and the population completely develops into female fishes and is sterile, so that the effect of obtaining all-female and sterile populations is achieved at the same time. The method provided by the invention has very strong applicability, and can be widely applied to aquaculture of fishes to obtain female unisexual groups of the fishes so as to improve the aquaculture yield, realize effective control on female fertility and protect ecological safety; the method has a wide application prospect in the fields of genetic breeding and ecological safety of aquaculture fishes.

Provided is a combination for specifically detecting P target genes. The combination comprises P pairs of primers used for amplifying P genes and P probes used for hybridizing the P genes. The P probes are respectively labelled with M different fluorescent groups having N concentrations; the types or concentrations of fluorescent groups carried by each probe among the P probes are different; and the P target genes and the fluorescent groups meet the following formula: 2≤P≤(MN+M)-X, M being a positive integer greater than or equal to 2, N ranging from 1 to 3, and X ranging from 0 to 19.

The invention discloses a corn crossbreeding intelligent matching system and method based on artificial intelligence, and the method comprises the steps: collecting molecular marker genotype data and phenotype data of candidate parents, calculating the genetic distance between the parents, and constructing a genetic distance matrix; analyzing the phenotypic complementarity degree of the parent pair, and calculating a phenotypic complementarity index; constructing a heterosis prediction model based on genetic distance and phenotype complementarity, and predicting the yield heterosis value of the candidate hybrid combination; evaluating the environmental adaptability and resistance comprehensive score of the candidate combination; and adopting a Pareto multi-objective optimization strategy to screen an optimal matching scheme. According to the method, molecular marker data and phenotype data are integrated, a heterosis prediction model considering a genetic distance nonlinear effect is constructed, collaborative optimization of multiple breeding targets such as yield, resistance and environmental adaptability is achieved, the prediction accuracy is improved by about 20% compared with that of a traditional method, and the method is suitable for large-scale popularization and application. And an intelligent and precise decision support tool is provided for corn crossbreeding.