173 results about "Library preparation" patented technology

The present disclosure provides methods for reducing sequencing errors comprising one or any combination of: (i) removing deaminated bases in any nucleic acid molecule throughout a library preparation workflow which includes immobilised splints which bind to the library, the use of a compaction oligonucleotide, optionally with an intervening sequence, formation of closed circular nucleic acids, creating gaps using glycosylase and lyase activities at positions with deaminated bases. The library may be sequenced using pairwise sequencing, e.g. with dark sequencing and / or sequencing using a multivalent labelled probe for the formation of an avidity molecule and soluble primer and polymerase. Method for sequencing concatemers in which the concatermers are contacted with polymerases, soluble primers and a multivalent labelled molecule which forms a complex with the polymerase. Detecting polymerase position and nucleobase bound to the polymerase in the complex. These methods generate higher quality base calls during downstream sequencing workflows.

The invention discloses a method and system for detecting respiratory tract pathogens, pathogen drug-resistant genes and pathogen virulence genes, and relates to the technical field of gene detection.The method comprises the steps that a respiratory tract sample of a patient is collected, DNA and RNA are extracted, and total nucleic acid is obtained; carrying out first-round PCR amplification on the total nucleic acid, purifying a first-round PCR product, carrying out second-round PCR amplification, and sorting the product to obtain a library building product; performing high-throughput sequencing on the library building product to obtain sequencing data, and detecting pathogens, drug-resistant genes and virulence genes; according to the method and system for detecting the respiratory tract pathogens, the pathogen drug-resistant genes and the pathogen virulence genes, by optimizing library preparation and direct library building of nucleic acid, the process is simplified, and efficiency and accuracy are improved; the balance reagent design guarantees library quality; the kit adapts to pathogen detection requirements in Yunnan and the like, provides efficient diagnosis, shortens detection time, improves treatment effect, regularly updates pathogen information, and keeps technical frontier.

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.

Described herein are methods, kits and systems for sample enrichment, multi-step library preparation, sample normalization, detection of sample biomolecules and combinations thereof. Enrichment and multi-step library preparation is described in the context of microfluidic workflows. Sample barcoding methods and kits are described for increasing sample throughput while reducing background in negative samples. Integrated microfluidic devices comprising sample processing unit cells coupled to an array of reaction sites are provided for integrated workflows.

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.

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

Disclosed herein are compositions and methods for detecting oligonucleotide molecules that can be ligated with high efficiency, and methods of using the oligonucleotides to tag DNA encoded libraries and to modify existing DNA encoded libraries to incorporate new functionalities. Also disclosed are compositions for increasing DNA solubility in non-aqueous solvents and assay systems for detecting compounds or conditions that increase the solubility or durability of DEL.

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.

The present disclosure provides compositions, including kits, comprising nucleic acid double-stranded splint adapters, and methods, e.g., PCR-free workflows, using double-stranded splint adapters. The double-stranded splint adapters (200) can be used in a one-pot multi-enzyme reaction to introduce one or more new adapter sequences into library molecules. The double-stranded splint adapters (200) comprise a first splint strand (long splint strand (300)) and a second splint strand (short splint strand (400)), which hybridize together to form the double-stranded splint adapter (200) having a double-stranded region and two adjacent single-stranded regions. The second splint strand (400) carries the new adapter sequence(s) being introduced, e.g., a universal binding sequence and / or an index sequence.

Compositions and methods are provided, for preparing DNA libraries for high- throughput sequencing following methylation-sensitive and / or methylation-dependent enzymatic digestion of the DNA. The provided compositions and methods obviate the need to clean-up the DNA sample between the digestion step and subsequent library preparation steps. The provided compositions and methods are particularly advantageous for library preparation from small amounts of DNA, such as cell-free DNA from body fluid samples.

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.

Methods are provided to capture telomere-associated DNA loci from chromatin. Chromatin was prepared from cultured cells by dual crosslinking using formaldehyde and EGS, followed by sonication. Biotinylated PNA probes were hybridized to the telomere DNA sequences, and serine protease was used to release the telomere-associated DNA sequences. The released DNA fragments in the supernatant were purified and subjected to library preparation for next-generation sequencing. The ligation-free chromatin capture and sequencing method enriches specific telomere-associated DNA loci to provide an accurate representation of chromatin interaction. Insights into the chromatin interactions occurring at telomeres are used to investigate the potential role of telomers in aging and age-related diseases.

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.

Massively parallel sequencing of cell-free maternal plasma DNA has recently been demonstrated to be a safe and effective method for screening for fetal chromosomal aneuploidy. Here, we report an improved sequencing method that significantly increases throughput and reduces costs by replacing the laborious sequencing library preparation step with PCR using a single primer pair. Using this method, samples containing as little as 4% trisomy 21 DNA can be readily distinguished from euploid samples.

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.

The invention relates to a fusion polypeptide containing a cyclic peptide with a novel configuration and a single domain antibody, a phage display library for displaying the fusion polypeptide, a preparation method of the phage display library, and application of the fusion polypeptide and the phage display library. The phage display library has the advantages that the display efficiency of the polypeptide is improved, meanwhile, the polypeptide can be prepared by using eukaryotic cell strain expression, and the purification of a polypeptide sample is realized through a single-domain antibody, so that the polypeptide with higher purity and lower immunogenicity is screened and obtained with low cost and low pollution.