97 results about "Target enrichment" patented technology

The presently claimed invention provides methods and kits for amplifying a target sequence from within a nucleic acid population. The presently claimed invention provides selection probes which are complementary to at least a portion of said target sequence and mechanisms for adding a probe sequence to the 3' end of a target sequence that is hybridized to a selection probe. The added 3' probe sequence and a probe sequence added at the 5' end of the target by adaptor ligation allow for selective amplification of the target sequence.< / PTEXT>

The present invention relates to a device for the specific selection of target molecules, comprising: (a) at least one reaction zone comprising a microarray, wherein the microarray comprises a substrate, on which one or more species of capture molecules are immobilized, comprising one or more temperature control and / or regulating units for controlling and / or regulating the temperature within the zone; (b) at least one non-reaction zone comprising one or more temperature control and / or regulating units for controlling and / or regulating the temperature within the zone, which is in fluid connection with the reaction zone; and (c) at least one transportation means capable of generating and / or regulating a fluid flow between said reaction zone (a) and said non-reaction zone comprising one or more temperature control and / or regulating units (b). The present invention further relates to a device for the specific selection of target molecules wherein the immobilized capture molecules are organized in the microarray in the form of spots, elongated spots and / or lines. In a further aspect the present invention relates to a method of specifically selecting target molecules, comprising the introducing a medium to such a device, performing interaction reactions in a reaction zone, transporting not interacted or not bound target molecules to a zone allowing reactivation of the target molecules and performing additional interaction reactions with the reactivated target molecules at the reaction zone, as well as the use of such a device for specifically selecting target molecules, e.g. for target enrichment also referred to as microarray based genome selection (MGS) in the literature.

The presently claimed invention provides methods and kits for amplifying a target sequence from within a nucleic acid population. The presently claimed invention provides selection probes which are complementary to at least a portion of said target sequence and mechanisms for adding a probe sequence to the 3' end of a target sequence that is hybridized to a selection probe. The added 3' probe sequence and a probe sequence added at the 5' end of the target by adaptor ligation allow for selective amplification of the target sequence.

The invention is directed to modified oligonucleotide compositions and methods for selectively reducing unwanted nucleic acid contaminants and enriching for desired nucleic acid targets from complex genomic nucleic acid mixtures for sequencing applications. The modified oligonucleotide compositions include one or more modified groups that increase the Tm of the resultant oligonucleotide composition.

The present invention relates to compositions and methods of target enrichment or selection of nucleic acids using hybridization, which can be used in, e.g., next-generation sequencing.

The present invention is directed to a method for reducing the complexity of a nucleic acid sample in a reproducible manner by enriching for specific nucleic acid target sequences in the population of nucleic acids. More specifically, the invention relates to a method for enriching specific target sequences in a population using libraries of oligonucleotides.

Provided herein is a capture library for target enrichment of sequences of interest from a genome DNA sample. The capture library comprises a plurality of capture oligos tiling a plurality of capture regions evenly-spaced along a genome. Each two adjacent capture regions of the plurality capture regions are separated by a spacing of about 6 to about 14 kilobases in length. The plurality of capture regions has a size of about 150 base pairs in length. Further, each capture oligo of the plurality of capture oligos comprises average 70 nucleotides in length. The capture libraries are suitable for enriching about 150 base pairs region approximately every 10 kilobases in a genome DNA. This capture library can be used to measure replication timing and copy number variation in human pediatric acute lymphocytic leukemia samples, and is also broadly applicable to any CNV application.

The present invention relates to sample-to-answer systems, devices, cartridges, and method of using the same for detecting the presence of microorganisms in a sample, such as bacteria.

The invention discloses a construction method of a genome presumptive area nucleic acid sequencing library and a device thereof. The method comprises the following steps: genomic DNA undergoes fragmentation treatment; DNA fragment undergoes fragment selection; the DNA fragment which has undergone fragment selection successively undergoes dephosphorylation, first end repairing and connection with first sequencing linker; a first connection product is screened by a probe; target fragment successively undergoes double-strands cyclization treatment and enzyme digestion; and an enzyme digestion product successively undergoes second end repairing, connection with second sequencing linker and DNA double-strands separation treatment so as to obtain single-stranded DNA which forms a genome presumptive area sequencing library, wherein the probe satisfies at least one condition selected from ten conditions. The probe has good specificity, high sensitivity and coverage in the aspect of target enrichment. By the method, the sequencing library for SNP enrichment detection can be effectively constructed.

The invention comprises methods and compositions for enriching for a target nucleic acid with a single primer extension and low-bias limited amplification.

The invention provides a construction method of a novel coronavirus whole-genome high-throughput sequencing library, and a kit for library construction. The method comprises the following steps: 1) carrying out reverse transcription on virus RNA; (2) carrying out a first round of PCR reaction by utilizing multiple amplification primers of an anchoring part Illumina linker sequence; and 3) carryingout a second round of PCR reaction by using a labeled Illumina library amplification primer, and purifying the amplification product to obtain the high-throughput sequencing library. The anchoring multiplex amplification primer combination provided by the invention is utilized, so the efficient targeted enrichment can be performed on the genome of the novel coronavirus COVID-19, the influence that an existing method is low in targeting, low in experimental timeliness and prone to being brought into host background pollution is overcome, COVID-19 virus whole genome sequencing can be completedwithin a short time with the small sequencing data volume and cost, so differential diagnosis and virus mutation identification of the COVID-19 virus are achieved.

The invention discloses an ultra-low frequency mutant nucleic acid fragment detection method, a mutant nucleic acid fragment library construction method, a primer design method and a reagent. Firstly,the fragmented nucleic acid of the sample to be tested is subjected to terminal repair, end-filling and adding A; connecting molecular tag joints; using the combination of molecular tag primers and single molecule specific primers to carry out amplicon targeted enrichment on the connection jointer product; finally, performing PCR amplification and enrichment. The single molecule specific primer designed by the invention obviously improves the number of available effective templates, and the carried adjustment sequence can obviously reduce the probability of forming primer dimers among specific primers, and the utilization rate of templates can be also obviously improved. The ultra-low frequency mutation nucleic acid fragment detection method of the invention uses one-sided single primer amplicon enrichment method and molecular label technology, improves the utilization rate of effective templates, reduces noise sources, reduces false positives, improves detection sensitivity and specificity, and is especially suitable for detection of trace ultra-low frequency mutation samples. At the same time, the invention can also carry out high-efficiency library construction.

The invention discloses a normal-pressure shale gas enrichment and high production target optimization method based on three-factor gas control. The normal-pressure shale gas enrichment and high production target optimization method comprises the following steps that (1) on the basis of single well dissection of a normal-pressure shale gas well and analysis of target enrichment and high production laws, the three main control factors affecting normal-pressure shale gas enrichment and high production are determined; (2) by taking the normal-pressure shale gas enrichment and high production main control factors as a main line, parameters highly related with the main control factors are optimized as analysis parameters; (3) different weights are given to the analysis parameters, and a normal-pressure shale gas enrichment and high production target optimization parameter list is established; and (4) an analysis target is subjected to quantitative classification analysis according to a target comprehensive evaluation value, and thus optimization of an exploitable area and well position deployment are guided. The normal-pressure shale gas enrichment and high production target optimization method has the beneficial effects that normal-pressure shale gas exploration of the Wufeng formation and the Long maxi formation in a southeast Chongqing district is guided effectively, and the exploration success rate is increased.

The invention provides a DNA homologous recombination anomaly detection method. The method comprises the following steps of (1) screening SNP sites; (2) designing a capture probe for the screened SNPsites; (3) carrying out genome DNA extraction and library construction; (4) carrying out library targeted enrichment; and (5) carrying out high-throughput sequencing, analyzing sequencing data, and using Kolmogorov-Smirnov test or scorHRD when the HRD state is judged.

The invention relates to an amplification primer group for detecting SARS-CoV-2 through mNGS and an application of the amplification primer group, and belongs to the technical field of gene detection.The amplification primer group comprises a random primer and an SARS-CoV-2 specific primer, the random primer is oligonucleotide of 5 bp to 7 bp, and the SARS-CoV-2 specific primer is oligonucleotideof 10 bp to 20 bp designed for an SARS-CoV-2 sequence. According to the amplification primer group, specific amplification is carried out on the novel coronavirus in a library building mode before library building, high-coverage targeted enrichment library building is carried out, and the coverage degree of pathogen detection, identification of pathogen genome mutation, the sensitivity of pathogen detection and the diagnosis capacity of novel coronavirus infection are improved.

Disclosed herein include systems, methods, compositions, and kits for performing targeted single cell mRNA sequencing assays. Capture oligonucleotides capable of specifically binding to target nucleic acid species are provided in some embodiments. Depletion oligonucleotides capable of specifically binding to undesirable nucleic acid species are provided in some embodiments. The method can comprise random priming and extension of barcoded transcripts followed by one or more enrichment and / or depletion steps using said capture oligonucleotides and / or depletion oligonucleotides. Immune repertoire profiling methods are also provided in some embodiments.

The invention discloses a drug-loaded magnetic microbubble, and a preparation method and application thereof. A drug-loaded nanometer particle packaged magnetic microbubble is formed by loading drug into mesoporous silica particles and self-assembling drug-loaded silicon dioxide nanometer particles and magnetic nanometer particles in a gas-liquid interface formed through a homogenizer in ice-bathenvironment. The drug-loaded magnetic microbubble is applied to drugs for treating phlebothrombosis, not only can protect thrombolytic drug, but also has rapid response, magnetic targeted enrichment and drug ultrasonic release, can increase the thrombolysis speed, and can solve low drug efficiency, toxic and side effect and bleeding complications in clinic thrombus treatment.

The invention discloses a polygene multi-target enrichment method. A capture probe library is designed, and target gene sequences are enriched in a probe capture way at a DNA (deoxyribonucleic acid) level, so that detection on all variation types, such as mutation, deletion, insertion, fusion and copy number amplification, of target genes is realized, and the defect that presently the target gene sequences are enriched by a PCR method at the DNA level, only mutation and deletion of the target genes can be detected, and the fusion cannot be detected is overcome.

A method of preparing a nucleic acid sample with target enrichment uses a reaction vessel (11), within which is added a chelating agent to a sample with heating to about 99° C. to provide a crude lysate. A PNA probe is provided at a concentration sufficient for binding and capture of discernible levels of target nucleic acid. The PNA probe may be attached to beads (26) which are initially embedded in a wax body (17) and are released during the heating so that they are free to move and come into contact with the PNA probe and target DNA. After binding has occurred, the beads are magnetically attracted back into a pocket (16) along with the wax (17), which is allowed to solidify before they are removed from the reaction vessel.

The invention discloses a CRISPR assisted DNA target enrichment method and an application thereof. sgRNA of a CRISPR / Cas9 system is reformed, novel sgRNA having an acquisition sequence at a 3' terminal is developed, the sgRNA and Cas9 protein without nuclease activity form a compound, the compound and target DNA are in target combination, and a formed DNA-dCas9-sgRNA compound can be acquired by magnetic beads of which the surfaces are fixed with single-chain acquired oligonucleotide, so that the target DNA is in target enrichment and separation from a DNA library or a mixture, and the CRISPR assisted DNA target enrichment method is used for sequence analysis of the target DNA. Compared with a target enrichment method based on hybridization, which is universal at present, the method provided by the invention has notable advantages of being high in simplicity, high in specificity, high in sensitivity, high in flux and the like, and can be deeply applied to preparation of reagents for DNAdetection, diagnosis and treatment.

The invention discloses an orthoester 5-fluorouracil prodrug molecule. The orthoester 5-fluorouracil prodrug molecule is N-5-fluorouracil1-ethyl-2-carbonalkoxy-(1,3) dioxolane-4-methylformamide havinga structure as shown in formula (I) described in the specification, wherein n=2-32 and m=n+2. The invention further discloses a preparation method of the abovementioned prodrug molecule and a nanoparticle prepared therefrom and an application thereof. The orthoester 5-fluorouracil prodrug molecule, the preparation method and the nanoparticle and the application thereof have the advantages that the nanoparticle prepared by the orthoester 5-fluorouracil prodrug molecule not only has the advantages of a small molecule prodrug but also is endowed with the advantage of a nano drug delivery system(that is a drug carrier), the prodrug nanoparticle not only has excellent acid degradation property and reduced tumor cytotoxicity and realizes passive targeting enrichment of a drug in a tumor, the nanoparticle has high drug loading capacity because the drug is used as a main part of the carrier, and the nanoparticle is capable of loading other antitumor drugs and achieves the effect of cooperatively treating the tumor and thus has good application prospects in the field of tumor treatment.

The present disclosure provides a method for enrichment of at least one target nucleic acid in a library of nucleic acids. A first oligonucleotide is hybridized to a target nucleic acid in library of nucleic acids having first and second adapters. The hybridized first oligonucleotide is extended with a first polymerase, thereby producing a first primer extension complex including the target nucleic acid and the extended first oligonucleotide. The first primer extension complex is captured, enriched relative to the library of nucleic acids, and a second oligonucleotide is hybridized to the target nucleic acid. The hybridized second oligonucleotide is extended with a second polymerase, thereby producing a second primer extension complex including the target nucleic acid and the extended second oligonucleotide, and further liberating the extended first oligonucleotide from the first primer extension complex.

The invention discloses applications of a fullerene structure in preparation of drugs used for treating leukemia. The fullerene structure is taken as an active ingredient, and comprises at least one ingredient selected from an oil soluble fullerene, an oil soluble metal embedded fullerene, a composition of the oil soluble fullerene and the oil soluble metal embedded fullerene, a water soluble fullerene, a water soluble metal embedded fullerene, a composition of the water soluble fullerene and the water soluble metal embedded fullerene, and a pharmaceutical ester or a pharmaceutical salt of theabove six ingredients. The active ingredient is capable of accumulating in bone marrow at a high efficiency; effects are achieved quickly; excellent biocompatibility is achieved; the fullerene structure can be used for preventing and treating leukemia before leukemia model maturation; after leukemia onset, the active ingredient possesses bone marrow part targeted enrichment characteristics, and can be used for rapid prevention, treatment, and repairing of bone marrow haematopoietic function damages by leukemia and other organic damages.

The invention provides a nanogel and a preparation method thereof. The method comprises the following steps: in the presence of a phenylboronic acid initiator and a morpholine initiator, carrying out reaction on L-glutamic acid oligo-polyethylene glycol monomethyl ether ester-N-anhydride in an organic solvent to obtain reaction liquid; and mixing the reaction liquid with L-cystine-anhydride and L-phenylalanine-N-anhydride and reacting to obtain the nanogel which comprises phenylboronic acid groups, morpholine groups, glutamic acid oligo-polyethylene glycol monomethyl ether ester chain links, phenylalanine chain links and cystine chain links and form a crosslinked network structure through disulfide bonds. The invention provides an anti-tumor nanogel drug loading system and a preparation method thereof. The drug loading particles have phenylboronic acid and morpholine groups on the surfaces to promote tumor targeting and endocytosis. A nanogel structure is formed through the disulfide bonds. After targeted enrichment and endocytosis in the tumor parts, the drug loading particles can quickly release the drug to achieve the effect of inhibiting tumors.