2336 results about "Genomic clone" patented technology

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.

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.

Multivalent binding compositions including a particle-nucleotide conjugate having a plurality of copies of a nucleotide attached to the particle are described. The multivalent binding compositions allow one to localize detectable signals to active regions of biochemical interaction, e.g., sites of protein-protein interaction, protein-nucleic acid interaction, nucleic acid hybridization, or enzymatic reaction, and can be used to identify sites of base incorporation in elongating nucleic acid chains during polymerase reactions and to provide improved base discrimination for sequencing and array based applications.

The invention belongs to the technical field of crop genetic breeding, and particularly relates to a rice hybrid male sterility site HMS10 co-segregation molecular marker and application thereof. The method comprises the following steps: hybridizing and backcrossing an anti-thermo-sensitive type genic male sterile line Yannong S serving as a female parent with a normal fertile material Yannong HB (serving as a recurrent male parent), finely positioning to a site HMS10 for controlling hybrid male sterility of rice in a physical interval of-60kb of a chromosome 10 of the rice by utilizing a high-generation backcross large population, and developing a co-separation molecular marker IDHMS10. The molecular marker IDHMS10 is utilized, through a molecular breeding strategy, a positive temperature-sensitive male genic male sterile line Quan 211S with excellent comprehensive agronomic traits is transformed into a practical rice hybrid male sterile line Quan 211HS with fertility not affected by light and temperature, and the IDHMS10 is used for purity identification of the hybrid male sterile line, so that the molecular marker IDHMS10 has great significance in solving the seed production risk of two-line hybrid rice and improving the yield of the hybrid male sterile line. And the method has important significance in enriching practical sterile resource types and the like.

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 invention discloses a method for targeted creation of a fast-growing new tilapia strain, and belongs to the technical field of fish cultivation. The method specifically comprises the following steps: (1) taking a fragment, as shown in SEQ ID NO.1, on a third exon of a pomc gene as a gRNA target sequence; (2) designing and synthesizing a gRNA primer sequence; (3) amplifying by adopting the gRNA primer sequence and the gRNA plasmid, and then transcribing and purifying to obtain a gRNA fragment; (4) transferring the gRNA fragments into fertilized eggs of tilapia mossambica through gene editing to obtain F0-generation positive fish; and (5) hybridizing the F0-generation positive male fish with the wild-type female fish, screening out F1-generation parent fish, and culturing the F1-generation parent fish to obtain F2-generation tilapia with homozygous deletion mutation. According to the invention, the pomc homozygous knockout line is established in the Nile tilapia mossambica for the first time, the new fast-growing line is a complete gene function deletion model, the cost is lower, the efficiency is higher, and the phenotype can be stably inherited.

The invention discloses an SNP (Single Nucleotide Polymorphism) molecular marker related to the daily gain character of a pig. The SNP molecular marker is a Ggt (Glutathione Ggt) located at the position of 160773437bp of a first chromosome of a pig genome Ensembl Sscrofa11.1 version; a mutation. Through the SNP molecular marker, the SNP molecular marker can be applied to breeding of the daily weight gain character of the Duroc long commercial pigs, so that the daily weight gain character of the Duroc long commercial pigs is improved, the growth performance and the slaughter weight of the Duroc long commercial pigs are improved, and the economic value and the economic benefit of the Duroc long commercial pigs are improved. And the variety of the Duroc long commercial pigs can be improved, the daily gain character of the variety is further improved, improved variety breeding is performed, and the market competitiveness of the Duroc long commercial pigs is improved. In addition, due to the fact that the Duroc long hybrid population is adopted as a research object, the Duroc long hybrid population can also serve as a reference population of a male parent Duroc purebred population, and reference and application values are improved for breeding of male parent Duroc boars in daily gain.

The invention provides application of a Zm00001eb378820 gene in molecular marker-assisted breeding for regulating and controlling the hundred-grain weight of corn grains, and relates to the technical field of agricultural biology. A backbone maize inbred line Ye107 is used as a male parent and hybridized with three excellent inbred lines (D39, R-2-1-1 and YML1218), a multi-parent group with remarkable hundred-grain weight difference is constructed, and SNP27712538 positioned on a chromosome 9 is a remarkable associated site by measuring hundred-grain weight phenotype data in multiple environments and combining high-density SNP (Single Nucleotide Polymorphism) markers, GWAS analysis and QTL (Quantitative Trait Loci) positioning. And the functional gene Zm00001eb378820 which is closely related to the regulation and control of the corn hundred-grain weight is excavated. A new insight is provided for a genetic regulation mechanism of the corn hundred-grain weight, and a key target spot is provided for breeding of high-yield corn varieties, so that the breeding efficiency is remarkably improved.

The invention relates to the technical field of high-throughput sequencing, and discloses a kit and a library construction method and a target variation detection and analysis method thereof, the kit comprises a pre-library construction reagent and a target region liquid phase hybridization capture reagent; wherein the pre-library construction reagent comprises a repair reagent, a joint, a connection reagent, an amplification reagent and purified magnetic beads; the target area liquid phase hybridization capture reagent comprises a hybridization reagent, a cleaning reagent, an amplification reagent, a purification magnetic bead, a target gene probe and a CNV / LOH probe. The invention provides a rapid, accurate and low-cost construction method of a library containing a targeting gene probe and a CNV / LOH probe and application of the library, through one-time library construction and sequencing, the advantages of a high-throughput targeting sequencing technology and a CNV-Seq technology are integrated; multi-gene point mutation, insertion and deletion, gene copy number variation, large-fragment copy number variation and heterozygosity deletion can be detected at the same time.

The present invention relates to ARP2 of plant stigma cells; the invention relates to application of Ca < 2 + > transport capacity damage of homologous genes 1 and 1 in breaking through reproduction isolation of plants. According to the method for breaking through the plant reproductive isolation, a foundation is laid for further research on the relationship of small peptides for regulating and controlling reproductive isolation calcium signals through co-receptors, and a simple and effective target is provided for gene modification for breaking through the reproductive isolation.

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 provides a tumor exosome (TDEs) fluorescence detection method based on tandem hybrid chain reaction (HCR) and selective coordination, and aims to combine an aptamer, a tandem enzyme-free amplification technology and selective coordination of Cu < 2 + > to analyze a clinical lung cancer blood sample. The programmed cell death ligand 1 (PD-L1) biomarker overexpressed on the surface of the TDEs is selected as a target for detecting the exosome. The HCR can easily generate a large amount of double-stranded DNA, so that the HCR can be used as a means for signal amplification. The high affinity of the aptamer to the protein will affect the tandem HCR reaction and then modulate the HCR product. Cu < 2 + > and a double-stranded DNA product are utilized to form a compound, and a fluorescence signal of calcein can be accurately adjusted, so that accurate fluorescence quantification is performed on the concentration of TDEs.

The invention relates to the technical field of protein interaction in yeast, in particular to a yeast double-expression vector, a construction method thereof, a protein interaction identification method and application. According to the vector, two exogenous genes can be cloned in the same vector, and the two exogenous genes can be expressed in yeast at the same time, so that interaction identification can be carried out. The work of plasmid construction and extraction, library preparation, yeast hybridization, screening culture and the like of an original yeast two-hybrid system is greatly reduced, and the identification efficiency of protein interaction is remarkably improved.

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 an SNP (Single Nucleotide Polymorphism) molecular marker combination and a primer group for identifying the authenticity of a mango 'Evin' and 'Jianghuang' hybrid and application of the SNP molecular marker combination and the primer group, and belongs to the technical field of mango crossbreeding. Comprising Mgchr < 111781329 > and Mgchr < 2081237 >; the Mgchr111781329 is located at the 1781329th site of a chromosome of chromosome11, the genotype of 'Evin' is TT, the genotype of 'Jinhuang' is AA, and the genotype of the hybrid is T / A; the Mgchr2081237 is located at the 81237th site of chromosome of chromosome20, the genotype of 'Evin' is AA, the genotype of 'Jinhuang' is TT, and the genotype of the hybridized F1 is A / T. A primer group designed on the basis of the SNP marker can quickly and effectively identify the authenticity of the hybrid of the mango 'Evin' and 'Jianghuang', the identification result is accurate and reliable, and genetic breeding of the mango is facilitated.

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.

Disclosed herein is a composition for nuclease treatment of hybridized probes in experiments to profile and analyze biological samples. Also disclosed herein, is a kit for nuclease treatment of hybridized probes in experiments to profile and analyze biological samples. Also disclosed herein, is a method for nuclease treatment of hybridized probes in experiments to profile and analyze biological samples.

The invention relates to the technical field of information, and provides a cross-platform global weed gene database unified building method which comprises the following steps: screening out a weed population having gene exchange characteristics with a target crop, identifying gene segments of gene exchange between the crop and weeds by adopting a gene comparison method, and determining a transmission path of resistance genes in interspecific hybridization; according to a gene comparison result, extracting transmission path data of the resistance gene, and analyzing the influence of resistance gene transmission on the genetic structure of the weed population in combination with the genetic structure characteristics of the weed population; calculating the frequency distribution of the resistance genes in different weed populations, judging the diffusion trend of the resistance genes in the weed populations according to the frequency distribution, generating a resistance gene diffusion map, and predicting the variation trend of the future weed population genetic structure based on a weed population genetic structure variation trend chart and the resistance gene diffusion map. And a global weed gene database is established, so that an effective weed management strategy can be formulated.

The invention relates to the technical field of medical biology, in particular to a method for identifying and diagnosing PEDV (Porcine Epidemic Diarrhea Virus) and PoRV-A. Colloidal gold is used as a carrier, and a monoclonal antibody (mAb) for resisting PEDV and PoRV-A and an initiator are respectively modified on colloidal gold (Au) to prepare a probe (Au-I-mAb). An ELISA hole is coated with a monoclonal antibody (mAb), when a target virus exists, a probe is added to form a double-antibody sandwich compound (mAb-antibody-mAb-Au-I), a hairpin is added, and an initiation chain on the compound recognizes the matched hairpin and induces the hairpin to cross to generate HCR. Due to the fact that the hairpins are marked by different fluorophores, when H1 is in a closed hairpin structure, fluorescence on H1 is quenched by a quenching group, after H1 is subjected to ring opening through HCR, the fluorophores are away from the quenching group, and fluorescence is recovered. Fluorescence excitation is carried out by using corresponding emission wavelengths, and two target pathogens are finally detected at the same time by detecting output signals of different fluorophores.

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.