133 results about "Library preparation" patented technology

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.

This document relates to compositions, kits, and methods for isolating target polynucleotides, provides oligomers, compositions, kits, and methods for capturing target polynucleotides, for example, for downstream applications such as amplification, library preparation, or sequencing. In some embodiments, a capture oligomer comprising a capture sequence that anneals to a complementary sequence is provided or used, the capture sequence can prevent capture until the complementary sequence is displaced in a target-polynucleotide dependent manner. In some embodiments, the amount of captured target polynucleotides is less than or equal to a predetermined amount.

The present disclosure provides compositions comprising nucleic acid double-stranded splint adaptors, including kits, and methods that employ the double-stranded splint adaptors, e.g., PCR-free workflows. The double-stranded splint adaptors (200) can be used in a one-pot, multi-enzyme reaction to introduce one or more new adaptor sequences into a library molecule. The double-stranded splint adaptor (200) comprises a first splint strand (long splint strand (300)) and a second splint strand (short splint strand (400)), where the first and second splint strands are hybridized together to form the double-stranded splint adaptor (200) having a double-stranded region and two flanking single-stranded regions. The second splint strand (400) carries the new adaptor sequence(s) to be introduced, such as for example a universal binding sequence and / or an index sequence.

The present disclosure generally relates to cartridge-based methods and devices for preparing a sequencing nucleic acid library from a sample. In embodiments, a cartridge-based method comprises placing a sample in a self-contained cartridge having a plurality of chambers, a reaction vessel, and a filter disposed in a fluidic path between the plurality of chambers and the reaction vessel, utilizing the filter to isolate the nucleic acid from the sample; forming a library prep reaction mixture; causing the library prep reaction mixture to flow into the reaction vessel and preparing the sequencing nucleic acid library comprising the tagged nucleic acid fragments, amplicons, and / or adapter labeled nucleic acid; isolating the tagged nucleic acid fragments, amplicons, and / or adapter labeled nucleic acid from the library prep reaction mixture on to the filter and eluting each from the filter; and purifying and optionally size selecting the sequencing nucleic acid library on the filter.

Provided herein are methods for generating circular nucleic acid molecules and circular nucleic acid libraries. The methods can be used to generate clonal populations of target nucleic acid molecules

The invention relates to the technical field of microfluidics and molecular diagnosis, and discloses a digital micro-fluidic chip for preparing an NGS methylation library. Comprising a bearing substrate, an electrode array used for electrowetting driving is formed on the upper surface of the bearing substrate to serve as a liquid drop conveying channel, the bearing substrate is divided into a plurality of functional areas which are sequentially arranged from the first side to the second side, the areas are communicated with one another through the liquid drop conveying channel, and micro-liquid drops are transferred and distributed among the areas. The upper cover is arranged opposite to the bearing substrate, a micro-droplet containing cavity is defined between the upper cover and the bearing substrate through a spacing frame arranged around the electrode array, and the upper cover is provided with a plurality of liquid inlet and outlet holes corresponding to the functional areas respectively. The chip is driven by the electrode array, so that micro-droplets entering the reagent sample introduction area and the sample introduction area through the liquid inlet and outlet holes are sequentially conveyed to the reaction area, and the micro-droplets after reaction are transferred to the taking-out area and are output through the liquid inlet and outlet holes, so that at least part of steps in the NGS methylation library preparation process are completed.

The invention provides a chain-specific library preparation method and a high-throughput sequencing method for rapidly detecting multiple types of RNAs (Ribonucleic Acid). The method comprises the following steps: artificially adding poly A tails at 3'tail ends of multiple RNAs by utilizing poly A polymerase; meanwhile, a polydeoxythymine ribonucleotide primer with deoxyuracil is used for synthesizing single-stranded cDNA under the action of reverse transcriptase, obtained single-stranded cDNA molecules are subjected to a series of reactions and finally subjected to PCR amplification to obtain strand specific libraries of multiple types of RNA, and samples of the libraries can be subjected to computer sequencing.

Provided herein are compounds of Formulae (I), (II), (III), and (III-d). Also provided herein are methods of preparation of Formulae (I), (II), and (III-d). Further provided herein are methods of functionalizing or sequencing a peptide by reaction of compounds of Formula (III-d) with peptidases.

Provided herein are compositions and methods for generating libraries for high throughput sequencing without using ligation, and methods of using same.

A library preparation device is a chip including (a) a biological sample well, (b) an end repair and elution well, downstream from the biological sample well, (c) an adapter ligation and elution well, downstream from the end repair and elution well, and (d) a final library well downstream from the ligation and elution well. The library preparation device is useful in a method of library preparation on a library preparation device.

The invention relates to compositions, kits and methods for isolating target polynucleotides, and provides oligomers, compositions, kits and methods for capturing target polynucleotides, for example for downstream applications such as amplification, library preparation or sequencing. In some embodiments, a capture oligomer comprising a capture sequence annealed with a complementary sequence is provided or used, the capture sequence preventing capture until the complementary sequence is substituted in a target-polynucleotide dependent manner. In some embodiments, the amount of target polynucleotide captured is less than or equal to a predetermined amount.

The use of different primer sets in reverse transcription incorporates tags allowing for the selective amplification of cDNA transcribed using the different primers. Primers targeting non-desired RNA sequences such as ribosomal RNA can be used to prevent subsequent amplification of cDNA transcribed from those non-coding fragments. Accordingly effective depletion of non-desired sequences after cDNA amplification can be achieved. Systems and methods of the invention have applications in whole-transcriptome analysis. Non-coding sequence targeting primers can include nucleotide analogs that, when enzymatically processed, prevent subsequent amplification. Library preparation can include single primer isothermal amplification (SPIA) techniques wherein an RNA sequence required for SPIA is included in random primers but is absent from primers targeting non-coding RNA.

Methods for controlling non-systematic error in an amplification-based next generation sequencing (NGS) library preparation are described, which method includes using an internal amplification control (IAC) sharing identical priming sites to a native nucleic acid target template of interest in a NGS library preparation.

1. Name of the product in this design: Packaging box (Free DNA Library Preparation Kit JKEasy). 2. Purpose of this design: As a packaging box. 3. The key design elements of this product are the combination of shape, pattern, and color. 4. The image or photograph that best illustrates the key design points: Design 1 3D view. 5. The design for which protection is sought includes color. 6. The bottom surface of this design product is a part that is not easily seen or not visible during use, so the bottom view of Design 1 is omitted; the bottom surface of this design product is a part that is not easily seen or not visible during use, so the bottom view of Design 2 is omitted. 7. Design 1 is designated as the basic design.

An automated nucleic acid processing system includes a plurality of independent, asynchronously operating processing bays, each receiving a single biological sample. Each bay contains a robotic liquid handling module performing addition-only liquid transfers, a thermal cycling module executing programmable temperature profiles, a magnetic bead separation module, and a gripper mechanism transporting sample vessels between modules. A controller independently controls each processing bay, initiates sample processing without batch accumulation, and dynamically schedules workflow steps across all bays. Each processing bay receives a single-use closed consumable cartridge containing pre-measured reagents for nucleic acid extraction, library preparation, CRISPR-mediated depletion, and magnetic bead-based normalization. The system generates a sequencing-ready nucleic acid library from each biological sample without intermediate fluorometric quantification of nucleic acid concentration. An output interface provides a normalized sequencing library directly loadable onto a high-throughput sequencing instrument.

1. Name of the design product: full-automatic library preparation system. 2. Use of the design product: nucleic acid library construction. 3. Design points of the design product: in shape. 4. Picture or photo best indicating the design points: perspective view.

Provided are DNA library preparation methods and compositions that duplicate a target nucleic acid sequence. A target DNA template including the target sequence is circularized via an end adapter to form a circular construct, which is bidirectionally extended by a polymerase-mediated extension that is initiated at nick sites of the end adapter. Following polymerase-mediated extension, a double-length DNA template is formed that includes two copies of the target DNA template (and hence two copies of the target sequence). Each strand of the double-length DNA template includes a parental polynucleotide strand joined to a newly synthesized daughter strand copy of the parental polynucleotide strand. Predetermined sequences can be included in the double-length DNA template, such a primer sequences, unique molecule identifiers, and sequence indexes. Sequencing of the double-length DNA template can reveal genetic / epigenetic information associated with the target sequence. Also provided are methods to create asymmetric and multi-length DNA template constructs.

The present disclosure relates, in general, to methods for improving preparation of a spatial transcriptomics RNA, library, for example a mRNA library, by improving capture of RNA transcript information from a tissue sample in situ. The spatial transcriptomics library from a tissue sample is useful to determine a genetic profile and help diagnose a person who has or is at risk of having a disease, such as cancer, genetic disease, autoimmune disease, and other indications, and improve treatment of the subject.

A method for detecting variants within one or more genes of interest, the method comprising: designing a set of capture probes to cover at least one of the one or more genes of interest, including coding regions and non-coding regions; using the set of capture probes to prepare a sequencing library using a hybrid capture target-enrichment library preparation workflow; and identifying intra-genic structural variants.

The invention provides a library building method for increasing the yield of a full-length target gene by coupling 10 * single cell sequencing. The library building method for increasing the yield of the full-length target gene by coupling a 10 * single cell sequencing system comprises a step of enriching a target gene template by asymmetric amplification and a step of amplifying a library obtained by nested amplification, wherein the full-length sequence template enrichment of the target gene is realized through asymmetric amplification, and then the full-length amplification of the high-fidelity target gene is realized through the nested amplification step. The invention also comprises a system for coupling 10 * single cell sequencing and increasing the yield of a full-length target gene. According to the method and the system, efficient, specific and full-length coverage amplification of the target gene is realized; the method is especially suitable for detection of low-abundance genes in a 10 * system.

The invention relates to a kit for extracting and sequencing mitochondrial genome DNA of small arthropods, a sequencing method and application of the kit. The kit comprises a reagent A; a reagent B; a reagent C; the invention discloses a Tn5 transposase system for preparing a sequencing library. Aiming at the particularity of small arthropods, the trace mitochondrial DNA extracted from a single individual of a small insect by a one-step method can be used for library preparation and high-throughput sequencing, and a simple, convenient, efficient and universal technology is provided for mitochondrial whole genome sequencing analysis of numerous types of small insects. The sequencing data volume required by mitochondrial genome assembly by adopting the kit or the method disclosed by the invention can be reduced by dozens of times or even thousands of times compared with the data volume of the existing total DNA method, the difficulty that mitochondrial genome sequencing analysis cannot be carried out by utilizing a single small insect at present is overcome, the waste of rare insect samples can be reduced, and the cost is reduced. And the sequencing cost of a single sample is greatly reduced.

The present disclosure provides for methods of preparing nucleic acid libraries for sequencing. In particular, the present disclosure is directed to a workflow which employs a first round of amplification utilizing a universal polymerase chain reaction approach instead of a platform specific indexed polymerase chain reaction approach. Applicant has unexpectedly discovered that a workflow employing a first round of amplification which utilizes a universal polymerase chain reaction approach increases duplex recovery and / or genomic equivalent recovery as compared with a workflow employing a first round of amplification utilizing a platform specific indexed polymerase chain reaction.

The disclosure relates to methods for detecting somatic variants in the DNA of a subject having or suspected of having a disease or disorder, such as cancer. In some embodiments, the method for detecting somatic variants comprises partitioning the sample into at least a hypermethylated subsample. In some embodiments, the methods further comprise performing error-correcting sequencing library preparation using DNA from the sample or hypermethylated subsample thereof. In some embodiments, the method for detecting somatic variants comprises high stringency identification of somatic variants. In some embodiments, the methods provided herein are used determine the presence of minimum residual disease.

The invention discloses a multiple myeloma sequencing method based on a targeted capture three-generation sequencing technology, a kit and application, and relates to the technical field of gene detection, and the method comprises the steps of sample treatment, targeted probe group design and targeted enrichment capture, three-generation sequencing library preparation, bioinformatics analysis and the like to identify and analyze three types of variations. According to the method, the defects that an existing layered system lacks an accurate genomics basis, and FISH and NGS technologies and combination strategies are low in flux, incomplete in coverage, incapable of synchronously detecting various variations, high in cost and complex in process are overcome, comprehensive information of three key variation types of multiple myeloma SNV, SV and CNV can be obtained at the same time through a single experiment and one-time sequencing, and the method is suitable for popularization and application. The detection efficiency is improved, and sample consumption, operation time and cost are reduced; all hot spot areas and translocation breakpoint areas of the most important clinical related genes of the MM are covered, and a gene variation map which is more comprehensive than that of a standard FISH Panel is provided.

The present disclosure relates, in general, to compositions for next generation sequencing and DNA library preparation, and systems and methods for use of the compositions in generating DNA libraries.

The present disclosure provides a method and composition for preparing a nucleic acid library.In some embodiments, the nucleic acid comprises cell-free DNA, including cfDNA that requires analysis, for example, by sequencing.The method can include restriction enzyme digestion, adapter ligation, and subsequent amplification, and can provide an improved approach to reduce the number of adapter-dimers generated during the process.In some aspects, the method for preparing a nucleic acid library can include: digesting DNA molecules with restriction enzymes to generate DNA fragments; ligating adapters to the DNA fragments by incubating with ligase to generate a mixture of adapter-ligated DNA fragments and adapter-dimers; amplifying the adapter-ligated DNA fragments to generate amplified adapter-ligated DNA fragments; and reducing the amount of adapter-dimers by distinguishing between the junction between adapter and DNA fragment and the junction between adapter and another adapter. TIFF2022530289000002.tif98128

The invention relates to a trace RNA acetylation library building and sequencing method, and belongs to the technical field of biology. According to the trace RNA acetylation library building and sequencing method provided by the invention, aiming at the problems of high RNA dosage and low sensitivity of the existing acRIP-seq technology, the initial RNA amount required by library building is greatly reduced to 500 ng level from conventional 50-500 [mu] g by optimizing the opportunity of removing rRNA in an acRIP-seq process (instead of removing rRNA in advance) and improving an impurity washing mode. According to the optimization, accurate detection of the ac4C modification level in the ng-level RNA sample is realized, and the application bottleneck of the technology in detection of rare cell subsets, clinical trace samples and ac4C low modification abundance RNA samples is effectively solved.

The present invention relates to a kit for extracting and sequencing mitochondrial genomic DNA of small arthropods, a sequencing method and the use thereof. The kit comprises a reagent A, a reagent B, a reagent C and a Tn5 transposase system for preparing a sequencing library. The provided kit or method is directed at the particularity of small arthropods and can achieve library preparation and high-throughput sequencing by using trace-amount mitochondrial DNA which is extracted from an individual small insect by a one-step method, thereby providing a simple and efficient universal technology for mitochondrial whole genome sequencing analysis of a wide variety of small insects. By using the provided kit or method, the volume of sequencing data required for mitochondrial genome assembly can be reduced by tens or even thousands folds than the volume of data of existing total DNA methods, thereby overcoming the problem of incapability to carry out mitochondrial genome sequencing analysis using individual small insects, and not only reducing the waste of precious insect specimens but also significantly reducing the sequencing cost for individual specimens.