718 results about "Genomic clone" patented technology

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 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.

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 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.

Methods for determining a location of a feature on a spatial array include (a) providing an array of features on a substrate, where a feature of the array includes a barcoded oligonucleotide having, in a 5′ to 3′ direction, a spatial barcode, a cleavage domain, and a constant sequence; (b) hybridizing a priming oligonucleotide to the constant sequence; (c) extending the priming oligonucleotide using the barcoded oligonucleotide as a template; and (d) determining all or a portion of a sequence of the extended priming oligonucleotide corresponding to the spatial barcode, or a complement thereof, and a location of the extended priming oligonucleotide, and using the location of the extended priming oligonucleotide to determine the location of the feature on the spatial array.

The invention discloses a high-specificity minimal residual disease (MRD) monitoring method and system based on circulating tumor DNA (ctDNA). The method comprises the following steps: receiving tumor tissue sequencing data of an UTUC patient, and generating a double-Panel target list containing personalized and fixed Panel; respectively extracting plasma cfDNA and leukocyte gDNA; performing vacuum concentration, hybrid capture and sequencing on the cfDNA library by using the double Panel lists, and performing deep sequencing on the leukocyte gDNA; constructing an individualized clonal hematopoietic mutation filtering database; actively filtering and rejecting clonal hematopoietic background mutation by utilizing a filtering database; and calculating an MRD load score based on the filtered tumor-derived mutation and outputting a report. The system comprises corresponding modules which are used for automatically executing the process. According to the invention, through cooperation of four major technologies of double-Panel design, process optimization, UMI error correction and active clonal hematopoietic filtration, MRD monitoring with extremely high sensitivity and specificity on UTUC is realized, false positive is significantly reduced, and the kit has drug resistance early warning potential.

The invention provides a macro virus group sequencing library construction method for removing host ribosome RNA through an enzyme method and a kit, and belongs to the technical field of biology. The method comprises the following steps: hybridizing total RNA (Ribonucleic Acid) of a sample with a DNA (Deoxyribose Nucleic Acid) probe aiming at host ribosome RNA to form a DNA / RNA hybrid chain; rNase H is used for digesting RNA in the hybrid chain, and DNase I is used for digesting the remaining DNA probe in the system; purifying to obtain virus RNA (Ribonucleic Acid); carrying out fragmentation treatment on the virus RNA; carrying out reverse transcription on the fragmented RNA to synthesize a first chain cDNA, then synthesizing a second chain cDNA, and carrying out terminal repair and dA tail addition to obtain double-chain cDNA; connecting the linker with the dT tail with the double-chain cDNA; and carrying out PCR amplification and purification on the connection product to obtain the macro virus group sequencing library. According to the construction method and the kit, the problem that the full-length sequence of the virus cannot be obtained from a low-load sample can be solved.

The invention discloses a citrus hybrid offspring authenticity identification method based on whole genome SNP (Single Nucleotide Polymorphism) analysis. According to the invention, a genome typing technology in a whole genome range is adopted, and a set of discrimination system capable of accurately identifying real hybrid offspring is established through high-density SNP (Single Nucleotide Polymorphism) marker analysis and an IBD (Identity by means of an algorithm. The method breaks through the limitation of a traditional method on hybrid filial generation identification, and provides reliable technical support for breeding practice.

The invention discloses a self-driven diagnosis and treatment integrated microneedle array paper-based chip as well as a preparation method and application thereof, and belongs to the technical field of biomedicine. The microneedle array chip system comprises a function integration module which at least comprises a self-powered extraction unit, a sensing detection unit and a controlled drug delivery unit. According to the microneedle array paper-based chip, biomarkers in skin interstitial fluid are efficiently collected based on a self-powered thermoelectric chemical battery microneedle, PtPB (at) PEG (at) HA nanoparticles loaded on an immunochromatography test strip combination pad are triggered to be subjected to ring opening, a hybridization chain reaction is induced to generate DNA nano-enzyme, and TMB is catalyzed to develop color; the open-loop nanoparticles can be used for SERS cross detection of biomarkers, and can be used as a thermal initiator to activate the phase change microneedle under the irradiation of near-infrared light, so that drugs are released as required, and precise treatment is realized. Besides, objective quantitative analysis of color signals is realized by constructing a convolutional neural network model, and the system has the functions of naked eye screening, intelligent quantification and personalized treatment, and can improve the home management ability of early skin cancer.

The invention relates to a male gamete killer gene S19g-A1 of a rice interspecific hybrid sterile S19 seat and application of the male gamete killer gene S19g-A1. The invention provides a rice hybrid sterility related protein S19g-A1. The amino acid sequence of the rice hybrid sterility related protein S19g-A1 is as shown in SEQ ID NO. 2. According to the invention, a key gene S19g-A1 of a rice interspecific hybrid sterile S19 site is researched and identified, function knockout is carried out on African rice S19g-A1, and pollen fertility of a mutant hybrid F1 obtained by carrying out distant hybridization on an obtained African rice s19g-a1 mutant returns to normal. Based on a gene editing technology, the invention develops a method for rapidly creating a hybrid affinity line capable of overcoming sterility of African rice hybrid, effectively eliminates S19 locus mediated interspecific hybrid reproductive disorder, breaks through the bottleneck of rice interspecific hybrid advantage utilization, provides target gene resources for distant hybrid advantage utilization and crop yield improvement, and has a wide application prospect. Good application prospects are realized.

The invention provides an invasive species identification method and device based on deep learning and DNA storage and electronic equipment, and relates to the technical field of invasive organism prevention and control, and the method comprises the steps: obtaining a to-be-identified invasive species image, and generating a first DNA sequence of a to-be-identified invasive species in the invasive species image through a deoxyribonucleic acid DNA encoder; obtaining a plurality of second DNA sequences to be hybridized from the invasive species database, and obtaining the hybridization yield of the first DNA sequence and each second DNA sequence by using a hybridization yield predictor; and determining the species category corresponding to the DNA sequence with the highest hybridization yield as the species category of the invasive species to be identified. According to the method provided by the invention, the strong feature extraction capability of deep learning is combined with the advantages of ultrahigh density and ultra-long stability of DNA as a data storage medium, so that end-to-end mapping and recognition from a species image to an exclusive DNA sequence thereof are realized.

The present disclosure provides methods of processing or analyzing a sample. A method for processing a sample may comprise hybridizing a probe molecule to a target region of a nucleic acid molecule (e.g., a ribonucleic acid (RNA) molecule), barcoding the probe-nucleic acid molecule complex, and performing extension, denaturation, and amplification processes. The nucleic acid molecule may be a nucleic acid molecule associated with CRISPR, (e.g., a guide RNA). A method for processing a sample may comprise hybridizing first and second probes to adjacent or non-adjacent target regions of a nucleic acid molecule (e.g., an RNA molecule such as a guide RNA molecule), linking the first and second probes to provide a probe-linked nucleic acid molecule, and barcoding the probe-linked nucleic acid molecule. One or more processes of the methods described herein may be performed within a partition, such as a droplet or well. One or more processes of the methods described herein may be performed on a cell, such as a permeabilized cell.

The invention discloses a composition, a detection method and a kit for SpaceAS variable splicing space in-situ detection of nucleic acid. The composition comprises a pi-type probe, a U-type probe, an O-type probe, an S-type probe and a signal probe, and the O-type probe adopts an in-vitro cyclization strategy, so that the problem of low hybridization efficiency caused by dependence on enzymatic cyclization reaction in a traditional hybridization method is effectively solved; meanwhile, an efficient probe hybridization system, a high-sensitivity signal amplification mechanism and a multi-round iterative imaging process are combined, so that the detection sensitivity and specificity are remarkably improved. The detection method has the advantages of high efficiency, high sensitivity, high specificity, good repeatability, strong expandability and the like through the synergistic effect of five-step single-stranded DNA probe hybridization reaction and one-step DNA amplification reaction, and can be widely applied to space in-situ detection and analysis of variable splicing events.

The utility model relates to a full-automatic all-in-one machine for NGS library building hybridization sequencing interpretation, which is used for sample adding and sequencing of a sequencing chip, and comprises a rack, a manipulator, a library preparation instrument and a sequencing device, the sequencing device comprises a gene sequencer, a sample adding clamping mechanism and a turnover mechanism, the library preparation instrument is used for sample adding of the sequencing chip, and the sample adding clamping mechanism is used for sample adding of the sequencing chip. The sample adding and clamping mechanism comprises a clamping tool and a clamping base, the clamping base is provided with a second positioning groove, the clamping tool is placed in the second positioning groove, the clamping tool clamps the sequencing chip, the overturning mechanism overturns the clamping tool to drive the sequencing chip for sample adding to overturn, the manipulator comprises an execution end and a clamping piece, the clamping piece is arranged at the execution end, and the clamping piece is arranged at the execution end. The clamping piece clamps the clamping tool, and the execution end moves among the library preparation instrument, the turnover mechanism and the gene sequencer, so that the sequencing chip is inserted into a preset position of the slot after being subjected to sample adding and turnover. The clamping tool clamps the sequencing chip, so that the sequencing chip is prevented from being overturned and falling off, automatic insertion of the sequencing chip is realized, and the sequencing efficiency is improved.

Disclosed are methods of preparing a composition of non-naturally occurring DNA, comprising: (a) extracting DNA from a sample of a subject; (b) contacting the extracted DNA or its derivative with a panel of oligonucleotide probes designed to hybridize to a plurality of preselected genomic regions, thereby generating selected DNA, wherein at least 50% of the preselected genomic regions are CpG regions that each comprises at least 3 CpG sites and a CpG density of at least 0.02 CpG / bp, wherein the preselected genomic regions cover 10-500 Mb of sequence space in human genome, wherein the extracted DNA or its derivative or the selected DNA or its derivative is further treated with an agent or a combination of agents that discriminates between methylated and unmethylated cytosines; and (c) performing high-throughput sequencing on the selected DNA or its derivative and generating sequencing reads.

This invention belongs to the field of molecular genetics and discloses a method and application for controlling fish fertility using in vivo gene editing technology. The applicant, for the first time, cloned a carp-specific U6 promoter and used this promoter to construct the U6 gene. MOVIE gRNA transgenic vectors and water The Cas9 vector allows for the establishment of an in vivo editing system. This strategy enables the heritability of sterility, a reproductive control trait, through hybridization of fertile parents to produce sterile offspring. The operation is highly efficient, simple, and environmentally friendly. Furthermore, since high-copy-rate short, scattered repetitive sequences exist in all fish species, this strategy has broad applicability across species.

The present invention relates to an artificial hybrid promoter, and more particularly, to an artificial hybrid promoter including an upstream activation sequence of GAL10 gene and a core promoter sequence of PGK gene, a recombinant expression vector including same, a host cell, and a method of producing a target protein using the host cell.

This invention relates to the fields of molecular biology and biomedicine, and discloses siRNA for the expression of the *Schistosoma japonicum* Hsc20 gene and its applications. The siRNA contains a nucleotide sequence that hybridizes to a target sequence of the *Schistosoma japonicum* Hsc20 gene, wherein the target sequence is selected from the sequences shown in SEQ ID NO. 1-SEQ ID NO. 4. The siRNA specifically inhibiting the expression of the *Schistosoma japonicum* Hsc20 gene provided by this invention can be used to interfere with the transcription and expression of the *Schistosoma japonicum* Hsc20 gene and the growth and development of *Schistosoma japonicum*. In vitro experiments have confirmed that the siRNA provided by this invention can efficiently silence the *SjHsc20* gene; and in vivo RNA interference experiments in mice show that this siRNA can induce 50.62% (…) P The insect reduction rate was <0.05% and 44.29% ( P A liver oocyte reduction rate of <0.01% is suitable for preparing drugs to treat schistosomiasis.

The invention belongs to the field of molecular breeding, and particularly discloses a molecular marker for identifying the width of tartary buckwheat seeds, a molecular marker primer group and application of the molecular marker, the SNP site of the molecular marker is located at the 22515278 bp position of the first chromosome of a tartary buckwheat reference genome, and the base polymorphism of the molecular marker is G / T; according to the invention, an F2 segregation population constructed by a wide-grain material KQ-1 and a narrow-grain material JQ-2 is subjected to whole genome re-sequencing and correlation analysis of grain width phenotype data, and an SNP locus which is extremely remarkably related to grain width is identified in a 22.51 Mbp interval of a first chromosome of tartary buckwheat; the genotype of the locus is highly consistent with the grain width phenotype, and the locus can be directly used for rapid identification of tartary buckwheat germplasm resources, early screening of filial generations and molecular marker-assisted breeding of high-yield tartary buckwheat varieties, and has important application value.

The presently disclosed subject matter relates to using a haploid inducing line (whether existing or created) and transforming the haploid line so that it encodes cellular machinery capable of editing genes. The transformed haploid inducing line is used as a parent in a cross between two plants. During pollination, the parental gametes fuse to form an embryo; and the gene editing machinery is also delivered to the embryo at this time. During embryonic development, one set of parental chromosomes are lost, and the gene editing machinery operates on the remaining set of chromosomes. Thus, at least one haploid progeny with edited genes is produced from the cross.

Described herein are compositions and methods for reducing adaptered-tag sequencing reads during the identification and nomination of on- and off-target CRISPR edited sites. One embodiment is a method for reducing adaptered-tag sequencing reads during the identification and nomination of on- and off-target CRISPR edited sites, the method comprising: contacting in an amplification reaction one or more adaptered-tag blocking oligonucleotides with an isolated genomic DNA having one or more tag sequences and adapter sequences; wherein the adaptered-tag blocking oligonucleotides comprise one or more blocking moieties and hybridize to adaptered-tag sequences at a junction region between the adapter and tag sequences to reduce amplification of the adaptered-tag sequences.

The present invention, among other things, provides technologies for detecting and / or quantifying nucleic acids in cells, tissues, organs or organisms. In some embodiments, through sequential barcoding, the present invention provides methods for high-throughput profiling of a large number of targets, such as transcripts and / or DNA loci.

The present application provides a mouse model for studying microsatellite stable and unstable intestinal cancer. Specifically, the present application provides the use of a hybrid mouse in a reagent or kit for preparing a mouse model animal for constructing MSS type intestinal cancer and MSI-H type intestinal cancer, wherein the mouse is a F1 generation of C57BL / 6 x BALB / c. The mouse model of the present application provides the same genetic background for MSS type intestinal cancer cells and MSI-H type intestinal cancer cells, making the research of MSS type intestinal cancer and MSI-H type intestinal cancer more scientific.

The invention discloses an oilseed rape gene BnaKNAT1s and an application thereof in creation of droop-type silique oilseed rape, and belongs to the technical field of plant genetic engineering. The gene comprises two copies of BnaA03.KNAT1 and BnaC03.KNAT1, and the CDS (Coding Sequence) sequences of the gene are respectively shown as SEQ ID NO: 1 and SEQ ID NO: 2. Targeted sgRNA is designed according to homologous regions of first exons of the two genes, nucleotide sequences of the sgRNA are shown as SEQ ID NO: 3 and SEQ ID NO: 4 respectively, CRISPR / Cas9 editing vectors are constructed, agrobacterium tumefaciens is transferred, and then rape is infected, so that a transgenic plant is obtained. And screening a homozygous mutant with two genes subjected to frameshift mutation and without a transgenic element, hybridizing the homozygous mutant with a good variety, and carrying out multi-round backcross and selfing to obtain a new germplasm of the sagittaria silique rape. According to the germplasm, the canopy structure is optimized, the light energy utilization rate and ventilation performance are improved, high-density planting is adapted, and gene resources and core materials are provided for high-yield breeding of oilseed rape.

The invention provides a molecular marker primer closely linked with a brown planthopper resistant major gene Bph48 of rice as well as a marking method and application of the molecular marker primer. A rice insect-resistant variety 75-9-3 (called C268 for short) and Yangdao 9 (called 9311 for short) are hybridized to obtain genotypes of F2 individual plants, genetic linkage analysis is performed in combination with the brown planthopper resistance level of an F3 family, a resistance major gene Bph48 carried by the insect-resistant variety C268 is identified, a Bph48 positioning region is reduced to a 370kb fragment limited by markers 1M30.093 and 1M30.471 on the two sides, and the Bph48 gene is identified to be a Bph48 gene. And the molecular marker is closely linked with the molecular marker 1M30.471. The molecular marker 1M30.471 disclosed by the invention can be used for effectively detecting whether an insect-resistant variety C268 and derived varieties (lines) thereof 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 Bph48 is obtained.

The application belongs to the field of fish development, and particularly relates to a molecular marker for identifying Nibea albiflora sperm-induced Pseudosciaena heteroclita gynogenesis offspring and hybrid offspring, a primer set thereof, a kit and application. The primer set for identifying the molecular marker of the Nibea albiflora sperm-induced Pseudosciaena heteroclita gynogenesis offspring and hybrid offspring comprises primers F and R, the nucleotide sequence of the primer F is shown as SEQ ID NO. 1, and the nucleotide sequence of the primer R is shown as SEQ ID NO. 2. Based on genomic data analysis, the application develops the molecular marker F / R which can effectively identify the Pseudosciaena heteroclita gynogenesis offspring and hybrid offspring in the juvenile stage. The method comprises extracting genomic DNA of parents and offspring, PCR amplification and agarose gel electrophoresis detection, and finally distinguishing the gynogenesis individuals and hybrid individuals by whether the Nibea albiflora parent marker is contained in the electrophoresis band.

The invention belongs to the technical field of crop breeding by a gene editing technology, and particularly relates to a method for creating a novel tomato cytoplasmic male sterility germplasm through gene editing and an application of the novel tomato cytoplasmic male sterility germplasm. In order to solve the problems that existing cytoplasmic male sterile materials in tomatoes are insufficient, production of tomato hybrid seeds is restricted and the like, gene editing is performed on MSH1 through a CRISPR / Cas9 technology, cytoplasmic male sterile single plants are identified in mutant selfing separation progenies and hybridized with wild types twice, and the tomato hybrid seeds are obtained. Finally, a novel non-transgenic tomato cytoplasmic male sterility material with the MSH1 gene recovered to a wild type is obtained, and meanwhile, the horticultural character of the original fertile material is kept unchanged.

The invention discloses a construction method of a conditional knock-down dynactin animal model of astrocytes and an application of the animal model of the conditional knock-down dynactin animal model of the astrocytes. According to the method, a Dctn1LoxP gene knock-in mouse is hybridized with an Aldh1l1-Cre / ERT2 transgenic mouse, a target genotype mouse is obtained through three rounds of breeding, 100mg / kg Tamoxifen is continuously injected into the intraperitoneal cavity of the 2-month-old mouse for 5 days, and specific knock-down of dynactin in brain and spinal astrocytes is realized, including knockout of p150Glue and reduction of DCTN4, p50 and Arp1alpha protein levels. The model has the advantages of being high in specificity, permanent in intervention aging and capable of covering multiple life stages, the defects of a traditional model are overcome, the model can be used for researching the influence of dynactin on astrocyte neurobiological functions, a reliable tool is provided for screening related targets of nervous system diseases, and the model has important application value.

Provided is a high-resolution promoter interaction fragment targeted capture method suitable for plants, which comprises: constructing a chromatin conformation capture pre-library; designing an RNA probe reversely complementary to a core promoter sequence; using the RNA probe to capture a promoter interaction fragment in the chromatin conformation capture pre-library; amplifying the library; and performing sequencing to obtain information of the promoter interaction fragment. By means of complementarily pairing hybridization between an RNA and a DNA, the capture method can specifically capture a promoter interaction fragment from a high-resolution chromatin conformation capture library, so as to obtain information of the promoter interaction fragment. Compared with existing capture technologies, the method exhibits greatly improved capture efficiency and has an obvious advantage in searching for distal regulatory elements.

The invention relates to genetic breeding, in particular to a method for cultivating kelp triploid by inducing a monosexual dihaploid (DH) gametophyte. The method specifically comprises the following steps: establishing a kelp DH sporophyte culture system, inducing DH sporophytes to generate unisexual DH gametophytes (2n) through spore-free propagation, and then hybridizing the unisexual DH gametophytes (2n) with haploid (n) gametophytes with opposite sex to obtain kelp triploid (3n) sporophytes. And determining the relative content of DNA in the nucleus of the hybrid sporophyte by using a flow cytometry, thereby identifying the chromosome ploidy of the hybrid sporophyte as 3n. The method can be used for cultivating new varieties of high-yield, high-quality, stress-resistant and sterile kelp, and has important significance for developing genetic breeding and new variety creation research in kelp and even kelp brown algae.