24results about How to "Lower Sequencing Costs" patented technology

The invention discloses a capture probe group for detecting pathogenic microorganisms and application of the capture probe group. The nucleic acid sequence of the capture probe group comprises a sequence as shown in SEQ ID NO. 1 to SEQ ID NO. 101. The capture probe group can cover and detect 119 types of bacteria, 32 types of fungi, 41 types of DNA viruses and 71 types of RNA viruses and covers common infectious disease pathogens, the kit containing the capture probe can cover and detect 88 types of drug-resistant and virulence genes, the detection performance of pathogenic microorganisms is greatly improved, and the sensitivity and specificity are improved.

The invention discloses a congenital agomphosis gene detection panel and application thereof, and belongs to the technical field of gene detection. The congenital odontia gene detection panel comprises probes for detecting or hybridizing 104 congenital odontia related genes shown in a capture table 1, a sample DNA and the panel are subjected to solution / chip hybridization, sequencing is carried out after DNA enrichment of a target area is completed, harmful sites or genes related to congenital odontia are screened / predicted according to a determination result, and the congenital odontia gene detection panel is used for detecting or hybridizing the 104 congenital odontia related genes. The expansibility, speed and resolution required for evaluating the target gene can be met, multiple genes in multiple samples can be evaluated at the same time, time is saved, and cost is reduced. Besides, the panel sequencing is high in coverage rate, strong in pertinence, high in detection sensitivity, higher in effectiveness, easier to manage and capable of more accurately, comprehensively and effectively detecting the related genes of the congenital odontoprisis and mutation conditions of the related genes, and accurate diagnosis and risk assessment of the congenital odontoprisis are achieved.

The invention belongs to the field of molecular biology and animal molecular breeding, and discloses a method for identifying genetic composition of a hybrid generation of megalobrama amblycephala and culter alburnus, a specific primer pool and application of the specific primer pool in identifying the genetic composition of the hybrid generation of megalobrama amblycephala and culter alburnus. According to the invention, qualitative and quantitative detection is carried out on the basic group condition of SNP sites on each pair of homologous chromosomes of megalobrama amblycephala and culter alburnus by innovatively utilizing a method of combining multiple PCR and high-throughput sequencing, so that an efficient identification technology is provided for genetic composition of filial generations obtained from other related species such as megalobrama amblycephala and culter alburnus. Compared with a whole genome sequencing gene, the method has the advantages that only a specific region on each chromosome is amplified and sequenced, so that the sequencing cost is effectively reduced, the analysis period is also shortened, and the method is an efficient technology for quickly identifying the genetic composition of a filial generation obtained by hybridizing sibling species such as megalobrama amblycephala and culter alburnus.

The invention belongs to the technical field of microbiological detection, and particularly relates to a primer group, a kit and a method for detecting a coxsackie virus group A type 6 virus whole genome and application. A set of specific primer group aiming at CV-A6 whole genome sequence amplification is designed, aiming at different CV-A6 strains (different regions and different separation time), the super-multiplex PCR amplification can be realized, the whole genome sequence is further captured, the pathogen capture can be quickly realized at low cost in disease outbreak and epidemic periods, and the primer group has good practical value.

This invention relates to a method for structural variant (SV) identification and genotyping in long-read family pedigree samples. The invention pertains to the field of vegetative variant (SV) detection in families, specifically focusing on methods for identifying and genotyping structural variants. The aim of this invention is to address the problems of existing family-based SV detection methods, which heavily rely on high-coverage sequencing, resulting in insufficient utilization of genetic characteristics and inaccurate SV detection and genotyping, as well as the high cost of sequencing multiple samples. This invention uses individual sequencing data from all family members as input, extracts variant features from each member, performs cluster analysis on the family feature set, assigns features to their respective members, and then uses three family feature signal correction methods to correct detection errors. Finally, SVs are located and anchored using Mendelian inheritance laws, and haplotype genotyping of SVs is completed using linkage information from long-read sequencing fragments.

The invention discloses a Marfan syndrome related gene capture probe group, a kit, a library construction method and application, and belongs to the technical field of gene detection. The sequence of the capture probe set is shown as SEQ ID No.1-149 and SEQ ID No.150-238, target genes FBN1 and FBN2 can be captured in a targeted mode, sequencing is carried out, the target genes are accurately sequenced, and the sequencing cost is reduced. The kit comprising the capture probe group can effectively capture the target gene, the detection cost can be reduced, and the detection sensitivity can be improved. According to the library construction method using the capture probe set, target genes are enriched through a sequence capture technology based on liquid phase hybridization, the target genes can be effectively enriched, the requirement for the sample size is reduced, and meanwhile the sequencing cost can be better controlled. The product and the method provided by the invention can be effectively applied to screening of related genes, and the related genes can be researched more deeply.

The invention discloses a solution mixing system and a solution mixing method.The solution mixing system comprises at least two reagent tubes, solution selection pieces and a flow cell, a liquid collection cavity and a liquid distribution cavity are formed in each solution selection piece, and each liquid distribution cavity is provided with a liquid inlet, a liquid outlet and a liquid collection opening; the liquid collecting opening is selectively communicated with any one of the liquid inlet and the liquid outlet, the liquid inlet is communicated with the reagent tube, and the liquid collecting opening is communicated with the liquid collecting cavity; the flow cell is communicated with the liquid outlet, and liquid in the at least two reagent tubes is suitable for flowing to the flow cell after passing through one or at least two solution selection pieces. According to the solution mixing system, the mixed solution is prepared at any time when used, the problem that the stability is poor due to the fact that the mixed solution is prepared in advance and placed for a long time is solved, different solutions can be prepared according to the actual situation, the solution preparation flexibility is high, the sequencing quality can be improved, and the sequencing cost can be reduced.

The present invention discloses a method for constructing a trace nucleic acid library. The method comprises the following steps: (1) obtaining a fragmented single-stranded DNA or cDNA that has been phosphorylated at the 5' end and hydroxylated at the 3' end; (2) performing 3' end auxiliary connection of the DNA fragments obtained in step (1) to extend into a double-stranded DNA; (3) connecting the 5' end of the double-stranded DNA obtained in step (2) to a linker, and then performing dual-template substrate amplification to obtain a library. The present invention improves the efficiency of preparing a trace library and reduces the purification steps, which is of great significance for constructing a library with a trace amount of DNA starting material.

The application belongs to the field of single cell sequencing, and discloses a method for constructing a microbial single cell genomics sequencing library, which comprises the following steps: 1) lysing microbial single cells; 2) adding an amplification reaction mixture to perform whole genome amplification; 3) adding a fragmentation reagent to obtain a fragmented amplification product; and 4) adding microbeads and an amplification reaction mixture to label the fragmented reaction product with a barcode, so as to obtain raw materials for constructing a microbial single cell whole genome sequencing library. The effective library molecules obtained by using the transposase complex linker sequence and the library construction method designed in the application can be compatible with an Illumina second-generation sequencer, and can be mixed with other sample libraries for sequencing, so that the sequencing cost is greatly reduced.

The present invention discloses a non-invasive early screening method and system for cancer based on the terminal sequence characteristics of cfDNA. The method uses a relatively low-depth whole-genome sequencing method to perform statistical analysis on the distribution frequency change trends of three consecutive groups of 3bp base combination categories upstream and downstream of the break position of cfDNA from tumors and cfDNA from healthy individuals, establish an early screening model for cancer, and achieve non-invasive early screening for cancer. The present invention draws an overall map based on the distribution of the types of combined DNA motif combinations in units of 3 bases for 9 consecutive positions at the 5' end, and uses the frequency as a training feature to establish an early diagnosis model for biliary and pancreatic tumors in the studied cohort, and evaluates the effectiveness of the model. This method uses a more accurate method of analyzing tumor DNA in the blood to find clues for early diagnosis of tumors, providing more solid and reliable data support for clinical precision applications.

The present invention provides a capture probe for a myeloid hematological tumor detection gene panel and a design method and a detection method thereof, belonging to the field of gene detection technology. The probe of the present invention includes a panel A DNA capture probe and a panel B cDNA capture probe; the panel A DNA capture probe includes a nucleotide sequence as shown in SEQ ID NO.1-2400, and the panel B cDNA capture probe includes a nucleotide sequence as shown in SEQ ID NO.2401-10559. The probe of the present invention can be used to detect all types of genes for myeloid hematological tumor detection, can solve the detection of myeloid hematological tumor gene mutation, copy number variation, fusion gene, and gene expression in one stop, can be used for detection at all stages of clinical practice, has strong portability, and can be applied to multiple detection platforms.

The invention belongs to the technical field of pathogen molecular diagnosis and high-throughput sequencing, and particularly relates to a novel bunyavirus whole genome capturing and sequencing method and kit for low virus load and high host background samples. Through the collaborative design of host nucleic acid reduction and two rounds of capture and enrichment, under the condition that the proportion of reads of an initial host source is greater than 90%, a high-coverage sequencing result of high-proportion base sites in L / M / S three fragments of the novel bunyavirus can still be obtained at a relatively low sequencing depth, the coverage integrity and the sequencing sensitivity of a whole genome are remarkably improved, and the method is suitable for large-scale popularization and application. And the sequencing cost and the result volatility are effectively reduced. Reliable whole genome data support can be provided for molecular typing, reassortment / recombination identification, aggregation epidemic situation analysis and transmission chain tracing of the novel bunyavirus, and the method has important clinical and public health application value.

The invention discloses a capture probe for detecting human immunodeficiency virus type 1, a plurality of capture probes are obtained by adopting imbricated design according to a plurality of reference genome whole-genome sequences of standard strains and subtype strains of HIV in an NCBI database, after a total RNA library is constructed, the capture probes are used for capturing the HIV whole-genome library, and the HIV whole-genome library is used for detecting the human immunodeficiency virus type 1. The probe sequence densely and repeatedly covers all areas of each reference genome, and the design parameters of the probe are as follows: the length is 120nt, and the imbrication frequency is 3X-6X Tiling. The invention further provides a kit comprising the capture probe and application of the kit. According to the capture probe, sequencing data is optimized, invalid data redundancy is reduced, sequencing efficiency is improved, and sequencing cost is reduced. Through whole genome sequencing, drug-resistant mutation and low-frequency mutation of a low-load sample can be detected, typing and virus load can be detected at the same time, the detection cost is remarkably reduced, and the detection efficiency is improved.

The present invention provides a selective inhibitory cyclization auxiliary sequence and application. The selective inhibitory cyclization auxiliary sequence is composed of an interfering sequence binding region a, a linker binding region a, a linker binding region b and an interfering sequence binding region b from the 5' end to the 3' end, and the 5' end of the interfering sequence binding region a has a phosphate group; the interfering sequence specifically bound to the 5' end of the interfering sequence binding region a is adjacent to the interfering sequence specifically bound to the 3' end of the interfering sequence binding region b, and the interfering sequence is an interfering sequence in a fragment to be sequenced; the linker binding region a and the linker binding region b are respectively used for specific binding to a double-end linker sequence. The problem that a large number of sequencing results for uninteresting sequences or interfering sequences exist in the sequencing data in the prior art can be solved, and the method is suitable for the field of high-throughput sequencing.

This invention discloses a computer program product and its application for data processing devices in the field of bioinformatics for ctDNA variant detection. The technical problem this invention aims to solve is how to detect ctDNA variants in early-stage cancer or postoperative minimal residual disease (MRD) under conditions of no UMI library construction and moderate sequencing depth. This invention constructs a set of supporting sequences for candidate variant sites, generating a sequence feature tensor and a fragment physical feature vector (including normalized fragment length). The former is input into a first neural network branch to extract sequence representation, and the latter into a second neural network branch to extract physical representation. A fusion module combines the sequence representation and physical representation, and a gating unit calculates the gating weight based on the physical representation and dynamically adjusts the contribution of the sequence representation, outputting the probability of the true ctDNA variant. This invention utilizes physical laws to suppress sequencing noise and can be applied to monitor MRD under conditions without molecular barcodes.

The application discloses a high-efficiency chromatin DNA binding map sequencing method. The method adopts BAMEA-Tn5 transposase. The BAMEA-Tn5 transposase is obtained by adding a specific nucleic acid aptamer sequence of a transcription factor to a 5' end of a linker sequence of Tn5 transposase, and the 5' end is provided with a modified group. The method is targeted to a transcription factor site in cells by the BAMEA-Tn5 transposase based on the recognition ability of the nucleic acid aptamer, and the BAMEA linker sequence and the tag are added at the site where chromatin DNA is combined with the transcription factor by the cleavage activity of the Tn5 enzyme. The sequencing method adopts the nucleic acid aptamer to target and recognize the transcription factor, greatly reduces the sequencing cost, and the nucleic acid aptamer has stable performance and strong reproducibility.

The present invention relates to a method for constructing a capture library suitable for next-generation sequencing platforms, comprising the following steps: 1) obtaining fragmented DNAs; (2) connecting the fragmented DNAs to a Y-shaped linker to obtain a pre-library; (3) hybridizing the pre-library with a hybridization probe to obtain a hybridization product; and (4) eluting the hybridization product to obtain a capture library. The present invention further relates to a kit for carrying out the above-mentioned method.

The present invention discloses a method for DNA molecular signal amplification and nucleic acid sequencing based on a solid phase carrier. The method comprises: fixing a carboxylated DNA primer on a solid phase carrier with amino groups through a condensation reaction, adding a circular DNA library and an amplification reagent to the solid phase carrier, and performing rolling circle amplification, wherein the DNA primer is combined with the circular DNA library through base complementation. The present invention fixes the DNA primer on a solid phase carrier, and utilizes rolling circle amplification to amplify the DNA into a linear spiral structure, thereby avoiding the accumulation of errors caused by bridge PCR amplification into clusters, improving sequencing accuracy, and also reducing the high duplicate ratio caused by bridge PCR, saving sequencing costs, and at the same time, performing isothermal DNA amplification, which has low equipment requirements, can reduce the burden on operators, and realize automated preparation.

The invention provides a probe, a primer, a kit, a method and application for human adenovirus whole genome sequencing. Aiming at a conserved sequence in an adenovirus genome, a plurality of pairs of primer probes are explored. According to the primer pair and the probe, the coverage degree of different adenovirus type genomes is greatly improved, and meanwhile, the amplification capacity of the primer aiming at targets with different lengths is enhanced. Eight adenovirus whole-genome sequences can be uniformly amplified, so that not only is the coverage degree of the whole-genome sequences improved, but also the capturing capacity for targets with different lengths is enhanced. In addition, the method has high tolerance for samples with different qualities, samples with low concentration or samples degraded to a certain degree can be subjected to high-uniformity total-sequence capture, and the method has high sensitivity and uniformity.

The present invention discloses a probe group, method and application for detecting pathogenic microorganisms, wherein the probe group specifically detects pathogenic microorganisms in a sample. In particular, in the field of central nervous system infection diseases, the defects of conventional central nervous system infection pathogen detection are overcome: pathogen targeted enrichment overcomes the defects of long cycle and low positive rate of traditional detection methods; overcomes the defects of high host ratio, low microbial load and low sensitivity of mNGS; targeted enrichment overcomes the low pathogen load of central nervous system infection itself and the difficulty of detecting some intracellular bacteria. The sensitivity and specificity of pathogen detection are greatly improved, and it has important clinical application value.

The embodiment of the invention discloses a gene sequencing method and a gene sequencing system. The gene sequencing method comprises the steps that a gene sequencing chip is provided, the gene sequencing chip comprises a plurality of micro-reaction chambers and a flow channel communicated with the micro-reaction chambers, one end of the flow channel is a sample inlet end, the other end of the flow channel is a sample outlet end, and the inner surface of the gene sequencing chip is subjected to hydrophobic modification; a sequencing buffer solution is introduced into the sample introduction end of the flow channel; introducing isolation gas into the sample introduction end of the flow channel to form an isolation gas column; introducing a water-phase reaction liquid into a sample introduction end of the flow channel; introducing sealing gas into the sample introduction end of the flow channel to form a sealing gas column; and carrying out a sequencing reaction through the gene sequencing chip. The gene sequencing method can at least partially solve the problem that in the prior art, the utilization rate of water-phase reaction liquid with high cost is low.

This invention belongs to the field of microbial detection technology and provides a high-throughput, low-cost targeted next-generation sequencing method, including sample collection and preprocessing, target gene amplification, direct library construction, multi-sample pooling, high-throughput sequencing, and data analysis. By optimizing the sample collection and preprocessing, target gene-specific amplification, direct library construction, multi-sample pooling, and high-throughput sequencing and data analysis processes, this invention significantly reduces sequencing costs and improves detection efficiency, while ensuring data accuracy and reliability. It effectively solves the problems of high cost, complex operation, large data volume requirements, and unstable results in existing technologies, and is particularly suitable for low-cost, multi-sample simultaneous detection needs in the field of microbial detection.

The application discloses a cancer non-invasive early screening method and system based on cfDNA sequencing coverage depth characteristics near TSS. The method realizes non-invasive early screening of cancer by establishing an early screening model of cancer through statistical difference of sequencing data coverage depth mode of tumor-derived cfDNA and cfDNA derived from healthy individuals near TSS through a low-depth whole genome sequencing method. The region of 500bp [‑250bp, 250bp] upstream and downstream of TSS is defined as a central region, and the upstream [‑2000bp,‑1000bp] and downstream [1000bp, 2000bp] of TSS are peripheral regions; the NF value of a gene is the average coverage of the central region divided by the average coverage of the peripheral region. The application adopts a low-depth whole genome sequencing method, greatly reduces the cost, and can detect abnormal changes of fragments earlier than cfDNA mutations in the early stage of cancer, and the method is more sensitive than detecting cfDNA mutation information.

The invention provides a multi-flow-channel space interval flow cell which comprises a flow channel defined by a first surface and a second surface and used for fluid to pass through. At least two flow channels exist, and each flow channel is provided with an independent liquid inlet and an independent liquid outlet; the first side faces of different flow channels are provided with physical space partitions. Meanwhile, the invention provides gene sequencing equipment which comprises a solution storage device which comprises at least one first reagent storage unit and is used for storing a sequencing biochemical reaction reagent; the fluid loading device comprises a first pipetting unit which can be used for synchronously and / or asynchronously loading various reagents in the solution storage device into the plurality of flow channels of the flow cell; and the power circulating device comprises a first recovery unit, is positioned between the flow cell and the solution storage device, and can isolate, collect and temporarily store the reagents flowing out of the flow cell according to different types, so that the reagents can be conveniently used in the subsequent sequencing process, and the effect of reusing the reagents is achieved.