83 results about "Hybridization capture" patented technology

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 belongs to the technical field of biochips, and particularly relates to a reusable genetic typing chip and a detection method thereof, the reusable genetic typing chip provided by the invention is prepared from polystyrene microspheres, capture probes and a silicon substrate, and depends on competitive hybridization of DNA in an SNP recognition stage, and the detection result is more accurate. The recognition sequence of the capture probe is converted into the sequence of the target object, and the added detection probes are hairpin probes with a self-protection function, so that the generation of false positive signals is remarkably reduced, and the detection specificity is remarkably improved. And no enzyme participates in the whole process, so that the detection process is remarkably simplified, and the detection cost is reduced.

Disclosed is a single-cell exon sequencing method and a use thereof, the method comprising: S1, tumor single cell sorting; S2, performing MDA amplification of sorted tumor single cells to obtain a genome library; S3, measuring the uniformity of MDA amplification products, selecting multiple sites located on different chromosomes and designing corresponding amplification primers, and testing the amplification effect of the multiple sites of the MDA product of each single cell by means of qPCR, thereby determining whether the genome has been uniformly amplified; S4, gDNA library preparation: breaking the amplified DNA into 150-200 bp fragments, and obtaining a gDNA library by means of end repair, tailing, adding adapters, and amplification; S5, exon capture and library construction: hybridizing the prepared gDNA library with a specific probe, capturing a specific region, and then adding an index tag via PCR amplification to obtain an exon library; and S6, sequencing the exon library.

The present disclosure provides a method for detecting the methylation and mutation status of a DNA sample, specifically, a method for simultaneously detecting the methylation and mutation status of DNA molecules in a sample, the method comprising treating the sample with a methylation-sensitive restriction enzyme, performing hybrid capture on amplification products of the restriction enzyme-treated sample and a sample that has not been subjected to enzyme cleavage treatment with a probe set including a mutation capture probe and a methylation capture probe, and sequencing the captured products.

The invention discloses a probe combination for detecting genetic susceptibility genes of prostatic cancer. The probe combination comprises at least one of sequences as shown in SEQ ID NO 1-89. The invention also discloses a kit comprising the probe combination and a method for detecting the genetic susceptibility gene of the prostatic cancer by using the kit. According to the present invention, the prostatic cancer genetic susceptibility related genes such as ATM, ATR, BRCA1, BRCA2, BRIP1, CHEK2, EPCAM, FANCA, HOXB13, MLH1, MRE11A, MSH2, MSH6, NBN, PALB2, PMS2, RAD51C, RAD51D and TP53 can be detected at one time by using the hybrid capture method; the probe provided by the invention is wide in coverage, high in sequencing depth and capable of detecting embryonic line variation of all exon regions of a target gene. All related diseases of the related genes of the genetic susceptibility of the prostatic cancer can be reported, genetic modes corresponding to the diseases are provided, and the application potential and the market value are large.

The invention provides a phosphorylated probe capture amplification method. The method comprises the following steps: hybridizing a phosphorylated probe to genome DNA to form a hybridization probe, extending and connecting the hybridization probe to form a closed loop by using a bridging primer to form a closed loop PCR product, and carrying out PCR amplification on the obtained PCR product to obtain an amplified product which is an enriched target area. According to the application, phosphorylation is carried out by taking the probe as a skeleton, a sequence on an Illumina Tru Seq joint and a bridged complementary sequence are added, by adopting a mode of increasing primer bridging, products after probe amplification are connected end to end to form cyclization, and by adopting a mode of increasing EXO1 / EXO3 enzyme digestion, the formation of a dimer is reduced, so that the detection sensitivity is improved, and the detection sensitivity is improved. According to the invention, small-volume hybridization and small-volume amplification modes are adopted, so that the molecular collision probability is increased. The method has ultrahigh specificity and accuracy, each target site is independently amplified, the PCR preference is avoided, the coverage uniformity is superior to that of multiple PCR and hybrid capture, the operation is simple and convenient, the period is short, and the experimental process only needs several hours.

A DNA methylation library construction method and a library obtained with same, a DNA hybridization capture method and a kit. The construction method comprises: providing a test sample comprising a plurality of double-stranded DNA fragments; degenerating the double-stranded DNA fragments into single-stranded DNA; carrying out ligation reaction on the single-stranded DNA and a double-stranded linker sequence to obtain single-stranded DNA linked with the double-stranded linker sequence; carrying out extension reaction on the single-stranded DNA linked with the double-stranded linker sequence to form an extended fragment, the extended fragment being linked with the double-stranded linker sequence of the single-stranded DNA; carrying out oxidation reaction on cytosine having methylation modification in double-stranded DNA linked with the double-stranded linker sequence to obtain double-stranded DNA having protected cytosine; removing the extended fragment in the double-stranded DNA having the protected cytosine to obtain single-stranded DNA having protected cytosine; carrying out deamination reaction on cytosine having no methylation modification in the single-stranded DNA having the protected cytosine to obtain single-stranded DNA having uracil; and carrying out amplification reaction on the single-stranded DNA having uracil to obtain a DNA methylation library.

The invention discloses a 20K liquid phase chip for precise identification and efficient breeding of local goose germplasm resources and application of the 20K liquid phase chip, and relates to the technical field of biology. The method comprises the following steps: S1, genome DNA breaking; S2, whole genome library construction; and S3, probe hybridization capture. By developing the 20K liquid phase chip, a plurality of genetic markers are integrated together in a high-density manner, the number of the markers is greatly increased, the coverage range of the markers is greatly expanded, efficient and comprehensive genetic analysis can be performed in the whole genome range, the problems of insufficient number of the markers and narrow coverage range in the prior art are solved, and the breeding accuracy is improved; the method can effectively improve the precise identification capability of local goose germplasm resources, optimize the breeding strategy and accelerate the local goose breeding process, thereby providing powerful technical support for genetic improvement and efficient breeding of local goose varieties.

Provided are a method and a combination for one-tube simultaneous detection of tumor-specific gene mutation and methylation, and the use thereof. The method comprises searching a database to select mutation and methylation sites of a cancer driver gene and a cancer suppressor gene, synthesizing capture probes correspondingly paired with a target interval, performing hybrid capture on an amplification product of a sample treated with a restriction endonuclease by using a probe set comprising tumor-specific gene mutation and methylation capture probes, constructing a high-throughput sequencing library containing a target region, and performing high-throughput sequencing to obtain the sequence of a target fragment, so as to obtain the results of methylation, point mutations and indel variation of a cancer-associated gene. The provided method enables one-tube simultaneous detection of mutation and methylation states of a cancer-associated gene in one assay, requires a low content of a gene to be detected, can be used for methylation and mutation detection of a tumor-specific DNA, and has low detection costs and high efficiency.

The invention provides a chlamydia whole genome detection method and application thereof, and further provides a probe for capturing a chlamydia whole genome, and the probe is designed according to a chlamydia genome sequence as a reference sequence; the probes are 3-10 layers of probes designed in an imbricated manner, the length of each probe is 90-120nt, and the total number of the probes is 50,000-200,000. A probe for capturing a chlamydia whole genome is used for performing hybrid capture and high-throughput sequencing analysis on a to-be-detected sample to obtain a whole genome sequence. According to the detection method developed by the invention, a whole genome sequence can be directly obtained from a clinical sample, the problem that the chlamydia is difficult to detect is solved, the detection efficiency and typing accuracy of the chlamydia are greatly improved, and the method has an important application prospect in clinical detection.

The present application relates to the technical field of gene detection, in particular to a SNP molecular marker for detecting donor cfDNA of a pregnant organ transplant patient, a detection method and device. The SNP molecular marker for detecting donor cfDNA of a pregnant organ transplant patient is shown in Table 10. The present application uses free DNA in peripheral blood of a pregnant patient and genomic DNA in peripheral blood of a biological father of a fetus as detection objects, adopts liquid phase hybridization capture and high-throughput sequencing technology, uses thousands of SNP sites, obtains the genotype of the fetus through the genotype of the couple, eliminates the influence of cfDNA from the fetus on the content of cfDNA from the transplanted organ, accurately determines the concentration of cfDNA from the transplanted organ, has higher detection sensitivity, can be detected multiple times only by using peripheral blood of the patient, and achieves the purpose of long-term dynamic monitoring.

The invention belongs to the field of medical detection, and particularly provides a solid tumor molecule residual focus detection method which comprises tissue baseline detection, probe design, peripheral blood MRD detection and result analysis and interpretation. According to the present invention, the technical steps such as optimized pre-library construction and hybridization capture process, customized probe design, unique fusion gene probe design and the like are improved, such that the problems of low detection sensitivity, low specificity, low accuracy and the like in the prior art are solved.

This application provides a method and system for infectious disease risk prediction based on a large language model and wastewater monitoring. The method includes: acquiring a multimodal time-series dataset of a target area in a first time period, wherein the multimodal time-series dataset includes wastewater epidemiological data sequences of the target disease, vaccination data sequences of the target disease, clinical monitoring data sequences of the target disease, environmental data sequences, and demographic data sequences obtained based on technologies such as multipathogen hybridization capture sequencing and PCR; converting the multimodal time-series dataset into a text time-series data sequence; inputting the text time-series data sequence into a preset risk prediction model to generate corresponding feature vectors, and predicting the trend of hospitalization rate changes of the target disease based on the feature vectors; and predicting the risk level of the target disease in the target area based on the trend of hospitalization rate changes and the wastewater epidemiological data sequence, thereby improving the accuracy of infectious disease area risk prediction.

Provided herein are systems, kits, compositions and methods for sequencing library preparation and sequencing workflow (e.g., for the identification of mutations). In certain embodiments, provides herein systems and methods to identically barcode both strands of templates, and PCR-based enrichment of each strand that does not require hybridization capture.

The invention discloses a library construction method for non-HPV (human papillomavirus) related cervical tumor and precancerous lesion molecular subtypes thereof based on high-throughput sequencing, which comprises the following steps: ultrasonically breaking genome DNA (deoxyribonucleic acid) of a detected sample to obtain a DNA fragment, modifying the tail end, connecting a joint, and performing purification and pre-amplification reaction, namely performing hybridization reaction on a pre-amplification sequence by using a hybridization capture reagent, according to the method, a hybridization capture reagent is adopted, DNA fragments combined through non-specific capture are enriched and removed, then an amplification reaction is carried out, a library is obtained, next-generation sequencing is carried out, original data are subjected to bioinformatics analysis, variation information is obtained, and the hybridization capture reagent comprises specific primers with the sequences shown as SEQ ID NO: 1-175. According to the present invention, the integration of the non-HPV-related cervical tumor and precancerous lesion molecule detection is achieved, the detection can be completed in one experiment, and the sequencing depth is improved by specifically enriching the target region so as to improve the sensitivity of the non-HPV-related cervical tumor and precancerous lesion molecule detection.

The invention discloses a mutation back noise filtering algorithm based on next-generation sequencing data, relates to the technical field of high-throughput sequencing data analysis, and aims to solve the problems that the prior art depends on a baseline, and is poor in adaptability and high in false positive. The algorithm comprises the following steps: preprocessing sequencing data to obtain a pileup file, classifying sites according to a sequence context, calculating an error rate, constructing a layered back noise model to check mutation, qualitatively determining real mutation and back noise by combining a sample type, and evaluating sample quality. The method does not need to construct a base line in advance, adapt to hybrid capture and amplicon sequencing, can improve mutation detection specificity and repeated sample consistency, guarantees result reliability, and is suitable for scenes such as tumor gene detection.

The invention discloses a preciseness-adjustable probe hybridization capture method which is characterized in that a hybridization resolution enhancing probe is added into a system of a probe hybridization capture target area, a potential non-specific capture area is competitively combined, and the hybridization capture resolution of the probe to the target area is improved. The binding strength of the hybridization resolution enhanced probe and a potential non-specific capture region is higher than that of the probe and the potential non-specific capture region, and the binding strength of the hybridization resolution enhanced probe and a target region is far lower than that of the probe and the target region, so that the detection sensitivity of the target region is not influenced. The method realizes local fine control of hybridization capture preciseness of the nucleic acid target area, and simultaneously achieves high-sensitivity and high-specificity detection of the nucleic acid target area compared with a traditional method.

The invention belongs to the technical field of genotyping, and particularly relates to a Tan sheep 50K liquid phase chip based on targeted capture sequencing and application thereof, the Tan sheep 50K liquid phase chip is composed of independently packaged probe mixed liquid and a hybridization capture reagent, the independently packaged probe mixed liquid comprises a Tan sheep 50K probe, and site information of the Tan sheep 50K probe relates to 50000 SNP sites. The prepared Tan sheep 50K liquid chip is mixed with a Tan sheep high-throughput sequencing library, a target site is captured and amplified and purified, and after a product is subjected to high-throughput sequencing, a sequencing result is compared with a sheep reference genome, so that the genetic typing of the Tan sheep to be detected is obtained. The method is applicable to resource evaluation, genetic identification and genome selection of the Tan sheep variety, can break through key technical evaluation restricting genome selection breeding, lays a foundation for building a Tan sheep genome selection sharing platform, and accelerates the realization of the Tan sheep from phenotype selection to genome selection.

The invention belongs to the technical field of gene sequencing, and particularly relates to a library building kit and probe for detecting blood tumor related fusion genes. According to the library building kit, only a trace amount of total RNA is needed to be used for cDNA synthesis, the initial quantity of a template can be as low as 10 ng, so that a cDNA library is built, then the library is subjected to hybrid capture and enrichment by using an autonomously designed probe hematologic tumor fusion gene Panel (RA Panel), the time consumption is short, the efficiency is high, and simultaneous building and on-machine sequencing of 384 libraries can be realized. According to the rapid hybridization system of the library building kit, the shortest hybridization time can reach 30 min, the size of a finally obtained library fragment is about 200-400 bp, and the rapid hybridization system can be compatible with various Illumina sequencing platforms for sequencing. According to the invention, through a simple and easy-to-use solution of NGS sequencing while synthesis, rapid and accurate typing of fusion genes which may affect blood tumor diagnosis is realized.

The invention belongs to the technical field of library construction, and particularly relates to a DNA targeted library construction kit and application thereof. The kit comprises a hybridization capture reagent combination, the hybridization capture reagent combination comprises a hybridization reaction solution, and the hybridization reaction solution comprises a hybridization reaction solution 1 and a hybridization reaction solution 2; the hybridization reaction solution 1 is prepared from betaine, tetramethylammonium chloride, Mg < 2 + > and Triton X-100; the hybridization reaction liquid 2 is prepared from formamide and N-methyl pyrrolidone; hybridization of the probe and the library is facilitated, the capture efficiency is superior to that of an existing hybridization reaction solution, and Panel with different sizes and different hybridization durations can be compatible; a small panel can also achieve relatively high and relatively stable capture efficiency, the hybridization time is selectable within 1-20 h, fast and slow hybridization reagents are unified, and production and simultaneous operation of multiple products are facilitated.

The present application belongs to the field of medical detection, and specifically provides a detection method for solid tumor molecular residual lesions, which comprises tissue baseline detection, probe design, peripheral blood MRD detection and result analysis interpretation. Through the improvement of technical steps such as optimized pre-library construction and hybridization capture process, customized probe design and unique fusion gene probe design, the present application solves the problems of low detection sensitivity, low specificity and low accuracy in the prior art.

Provided herein are systems, kits, compositions and methods for sequencing library preparation and sequencing workflow (e.g., for the identification of mutations). In certain embodiments, provides herein systems and methods to identically barcode both strands of templates, and PCR-based enrichment of each strand that does not require hybridization capture.

The invention belongs to the technical field of hereditary deafness molecular diagnosis, and particularly relates to a deafness gene therapy related gene capture probe set, a detection method and application, the capture probe set comprises any one nucleotide sequence in SEQ ID NO.1-SEQ ID NO.404; the detection method comprises the following steps: carrying out hybrid capture on a nucleic acid sample to be detected by using the capture probe group; constructing a library for the captured target sequence, and performing high-throughput double-end sequencing; comparing the sequencing result with a reference genome, and identifying mutation of genes related to deafness gene therapy; the invention also discloses application of the capture probe group for the deafness gene therapy related genes in preparation of detection products for deafness genetic screening, deafness cause diagnosis, deafness genetic typing or gene therapy decision. Compared with the prior art, the gene capture probe set for deafness related to gene therapy is designed, the problem of non-uniform coverage of conventional sequencing in such areas is solved, and the whole genome capture uniformity and efficiency are improved.

The invention belongs to the technical field of molecular detection, and discloses a nucleic acid hybridization capture immunofluorescence detection kit as well as a use method and application thereof. In the kit provided by the invention, guanidine hydrochloride, guanidine isothiocyanate, triton X-100, NaCl, protease K and lauryl sodium sulfate are compounded according to a specific concentration to obtain the nucleic acid release liquid, so that pathogen nucleic acid in a clinical sample can be fully dissociated, and the nucleic acid release liquid can cooperate with a probe aiming at a specific target pathogen; the solution system can be used for optimizing the recognition and combination process of DNA and RNA, the specific recognition and combination capacity between the probe and specific pathogen nucleotide is improved, and under the condition that a common nucleic acid stabilizer, namely Tween, is not contained, the solution system can further strengthen differential structural epitopes existing in a DNA / RNA complex compared with DNA and RNA, so that the specificity of the DNA / RNA complex is improved, and the specificity of the DNA / RNA complex is improved. The kit can be better recognized and combined by an anti-DNA / RNA complex antibody, and has excellent detection specificity, sensitivity and precision.

In order to solve the willow breeding problem with low cost, high efficiency and simple means, the willow whole genome 40K SNP liquid phase chip is provided, the willow whole genome 40K SNP liquid phase chip comprises a willow 40K site probe mixed solution and a hybridization capture reagent, the willow 40K site probe mixed solution comprises a willow SNP background site probe and a willow SNP associated site probe, and each site comprises at least two capture probes. The liquid-phase chip is used for willow genetic diversity analysis, molecular genetic map construction, whole genome association analysis, variety authenticity identification, molecular marker-assisted selective breeding and whole genome selective breeding, is applied to willow leaf detection tests, performs statistics on detection conditions, has 40188 loci, and can be used for detecting willow leaves. According to the method, the average detection rate of the original SNP sites of 16 samples is 97.39%, the site detection rate reaches 99.9% after data of the 16 samples are merged and counted, the service cycle of sample detection is greatly shortened, and the breeding efficiency of willows is improved.

This invention relates to the field of pathogen detection technology, specifically to an intelligent adaptive virus detection and optimization method and system. The method includes: acquiring raw sequence data generated by probe capture sequencing; performing real-time analysis of the sequence data using a deep learning model, outputting analysis results including the virus's presence state and sequence characteristics; dynamically adjusting control parameters of the hybridization capture process based on the analysis results through an adaptive control system; and completing hybridization capture according to the adjusted parameters and outputting a detection report. The system includes a sequencing data acquisition module, an analysis module, a sensor data acquisition module, an adaptive control module, and a detection report generation module. This invention, by combining artificial intelligence analysis with real-time process control, achieves intelligent optimization of the virus detection process, significantly improving the accuracy, sensitivity, and adaptability of detection. It is particularly suitable for detecting highly variable viruses and low-viral-load samples, and has significant clinical application value.

The invention belongs to the technical field of gene sequencing, and particularly relates to a library building kit and a probe for detecting hematologic tumor chemotherapy related gene polymorphism by using a hybrid capture method. According to the present invention, the genotype of the hematologic tumor chemotherapy related gene, the drug treatment effect, the toxicity and the metabolic capability related information can be rapidly obtained through the one-time detection; meanwhile, the library building kit is simple and convenient to operate and short in consumed time, and automatic detection is extremely easy to realize. The detection result completed by using the library building kit is accurate, and the library building kit has important reference significance for treatment and prognosis of hematologic tumor patients needing chemotherapy.

The present disclosure provides systems and methods for targeted indirect, synergistic hybridization capture of a template for amplification and analysis of target sequences. The captured templates ca

An embodiment of the present disclosure provides a digital microfluidic chip, comprising a substrate and a cover plate. A ground electrode and a plurality of control electrodes are provided on the substrate, and a dielectric layer and a first hydrophobic layer are sequentially coated on the plurality of control electrodes. The plurality of control electrodes are used to drive the movement of reagents so as to perform methylation conversion, pre-library preparation, hybridization capture and final library preparation for the sample to be tested. The cover plate covers and is connected to the substrate, and sample injection holes for injecting reagents are provided at positions on the cover plate corresponding to at least a part of the plurality of control electrodes. A conductive layer and a second hydrophobic layer are sequentially coated on the side of the cover plate facing the substrate, and a gap for the movement of reagents is formed between the second hydrophobic layer on the cover plate and the first hydrophobic layer on the plurality of control electrodes, and the conductive layer is electrically connected to the ground electrode through a conductive glue.

This invention provides a DNA hybridization capture device with a positioning component. The DNA hybridization capture device with a positioning component includes: a rectangular column; sliding components are installed on the outer walls of the left and right sides of the rectangular column; the sliding components are connected to a supporting component; the supporting component is connected to a clamping and positioning component; and the clamping and positioning component is connected to the capture device body. The DNA hybridization capture device with a positioning component provided by this invention allows the rotating shaft to rotate via a worm gear, which in turn drives a worm wheel to rotate. The worm wheel then moves a rectangular plate along a rack, allowing the capture device body to be moved to a suitable height to accommodate workers of different heights. The worm gear and worm wheel also provide a self-locking function when the handle is not rotated. When the capture device body is placed on the supporting plate, the output of the motor drives a bidirectional threaded rod to rotate, causing two sets of threaded blocks to simultaneously move the clamping plate towards the outer wall of the capture device body until the capture device body is stably clamped.