311 results about "Library preparation" patented technology

The invention generally relates to sequencing library preparation methods. In certain embodiments, two or more template nucleic acids are joined together by a linking molecule, such as a PEG derivative. Identical copies of a nucleic acid fragment or both strands of a duplex fragment may be linked together. The linked nucleic acids are amplified, creating linked amplicons. Emulsion PCR with linked primers creates linked template nucleic acids for seeding sequencing clusters and errors can be readily identified by their presence on only one of the linked fragments.

The invention generally relates to sequencing library preparation methods. In certain embodiments, two or more template nucleic acids are joined together by a linking molecule, such as a PEG derivative. Identical copies of a nucleic acid fragment or both strands of a duplex fragment may be linked together. The linked nucleic acids are amplified, creating linked amplicons. Emulsion PCR with linked primers creates linked template nucleic acids for seeding sequencing clusters and errors can be readily identified by their presence on only one of the linked fragments.

There is described herein, a method of capturing cell-free methylated DNA from a sample having less than 100 mg of cell-free DNA, comprising the steps of: subjecting the sample to library preparation to permit subsequent sequencing of the cell-free methylated DNA; adding a first amount of filler DNA to the sample, wherein at least a portion of the filler DNA is methylated; denaturing the sample; and capturing cell-free methylated DNA using a binder selective for methylated polynucleotides.

This is described herein, a method of capturing cell-free methylated DNA from a sample having less than 100 mg of cell-free DNA, comprising the steps of: subjecting the sample to library preparation to permit subsequent sequencing of the cell-free methylated DNA; adding a first amount of filler DNA to the sample, wherein at least a portion of the filler DNA is methylated; denaturing the sample; and capturing cell-free methylated DNA using a binder selective for methylated polynucleotides.

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 relates to the technical field of protein interaction in yeast, in particular to a yeast double-expression vector, a construction method thereof, a protein interaction identification method and application. According to the vector, two exogenous genes can be cloned in the same vector, and the two exogenous genes can be expressed in yeast at the same time, so that interaction identification can be carried out. The work of plasmid construction and extraction, library preparation, yeast hybridization, screening culture and the like of an original yeast two-hybrid system is greatly reduced, and the identification efficiency of protein interaction is remarkably improved.

The invention relates to the technical field of biological extraction. The invention discloses an automatic control method and device for nucleic acid extraction and library preparation and a storage medium. The method comprises the following steps: acquiring to-be-analyzed process card information; predicting process card prediction information corresponding to nucleic acid extraction and library preparation through the target analytical model; generating a target process flow and target material information from the to-be-analyzed process card information through the process card prediction information; the target technological process is packaged in a segment program to generate a segment program combination file, and the segment program combination file is transmitted to a target module through a bus; and controlling the target module to automatically perform nucleic acid extraction operation or library preparation operation according to the segment program combination file. According to the method, the technological process can be automatically generated, and the accuracy of nucleic acid extraction and library preparation in automatic control is improved.

The present invention provides a preparation method for an extrachromosomal circular DNA library, a sequencing method, a kit, and a use thereof. The preparation method comprises: extracting total DNA in a sample and digesting linear DNA in the total DNA to obtain a digestion product, the digestion product comprising extrachromosomal circular DNA; directly carrying out rolling circle amplification on the digestion product without purification, to obtain an amplified product; and directly carrying out library preparation on the amplified product without purification, to obtain an extrachromosomal circular DNA library.

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.

Aptamer detection techniques are provided that facilitate sequencing library preparation. In embodiments, reporter probes complementary to aptamers may be used to generate amplification products to form a sequencing library. In embodiments, the aptamer may be used as part of a library preparation.

The present disclosure relates generally to methods for improving the preparation of spatial transcriptomic RNA libraries, such as mRNA libraries, by improving in situ capture of RNA transcript information from tissue samples. The spatial transcriptomic library from a tissue sample can be used to determine genetic maps and help diagnose persons having or at risk of having diseases, such as cancer, genetic diseases, autoimmune diseases, and other indications, and improve treatment of subjects.

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.

The invention relates to a tandem repeat related gene mutation detection system and a non-disease diagnosis purpose method. The system comprises an acquisition module, a Y-shaped pre-library preparation module, a targeted enrichment module, a single molecule real-time sequencing library construction module and a single molecule real-time sequencing analysis module, wherein target genes comprise ATXN1, ATXN3, ZFHX3, CACNA1A, ATXN7, ATXN8OS, ATXN10, PPP2R2B, TBP, FGF14, BEAN1-TK2, NOP56, DAB1, THAP11, FXN, GLS, RFC1, LRP12, GIPC1, RILPL1, VWA1, AR and DMPK; detection is carried out through three-generation sequencing, mutation setting is accurate, and the problem that an existing detection scheme is not practical is solved.

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.

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 invention provides a preparation method of a chromosome outer circular DNA library, a sequencing method, a kit and application thereof. The preparation method comprises the steps that total DNA in a sample is extracted, linear DNA in the total DNA is digested, a digestion product is obtained, and the digestion product comprises chromosome outer circular DNA; directly carrying out rolling circle amplification on the digestion product without purification to obtain an amplification product; an amplification product is directly subjected to library preparation without being purified, and the chromosome outer circular DNA library is obtained.

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.

To provide a practical method for detecting repeat expansion diseases that can comprehensively test for repeat expansion diseases without the need for advanced specialized knowledge in repeat expansion diseases.SOLUTION: The method for detecting a repeat expansion disease according to the present invention includes a library preparation process, a sequencing process, a consensus sequence construction process, a ranking process, and a determination process. In the sequencing process, multiple disease-associated repeat regions and their neighboring genomic regions are designated as target regions, and nanopore sequencing is performed to obtain long-read sequence data for each target region in a subject's genome. In the ranking process, repeat number data for each disease-associated repeat region is compared between a subject and a group of normal healthy persons, and the disease-associated repeat regions in the subject's genome are ranked in the descending order of the possibility of pathological repeat expansion. In the determination process, evaluation is started from the disease-associated repeat region ranked first according to specific steps S1 to S7, and it is determined whether or not the subject has pathological repeat expansion.SELECTED DRAWING: Figure 1

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.

The invention discloses an MMLV reverse transcriptase mutant. The invention provides an MMLV reverse transcriptase mutant, and compared with a wild type MMLV reverse transcriptase amino acid sequence, the mutant mutates at a plurality of sites such as a 62 site and the like, and the mutant has the characteristics that the template conversion performance is improved, the non-template basic group addition performance is improved or the heat resistance is improved. The MMLV reverse transcriptase mutant with improved conversion performance compared with a wild type template is screened out, and the MMLV reverse transcriptase mutant also has improved non-template base addition performance or thermal stability; the screened mutant is suitable for RNA (Ribonucleic Acid) sequencing, such as Smart-Seq, nanopore sequencing, 5 'RACE (Rapid Amplification of cDNA Ends) and the like, adopting a library preparation method based on template conversion, and can also be applied to RT-qPCR (Reverse Transcription-Quantitative Polymerase Chain Reaction) and the like.

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

There is described herein, a method of capturing and analyzing cell-free methylated DNA in a sample. The method involves subjecting the sample to library preparation to permit subsequent sequencing of the cell-free methylated DNA. A predetermined amount of control synthetic DNA fragments are added to the sample. The control synthetic DNA fragments each have a known nucleic acid sequence that does not align to a target genome sequence, and at least some of the control synthetic DNA fragments are methylated. The sample is denatured, and cell-free methylated DNA and the control synthetic DNA fragments are captured using a binder selective for methylated polynucleotides. The captured DNA is amplified and sequenced.

The invention relates to the technical field of methylation sequencing, in particular to the technical field of methylation library preparation of trace or degraded genome DNA. The invention also relates to a kit for microscale or degradation of methylated libraries of genomic DNA. The method and the kit provided by the invention are particularly suitable for sequencing analysis of a methylation target area, and the method and the kit are suitable for library construction of trace or seriously degradable samples.

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.

The invention provides a method for screening an aptamer from an RNA (Ribonucleic Acid) library. According to the method, the eluent of each elution is collected, and meanwhile, a specific elution procedure is combined, so that the technical effects that any information of the RNA aptamer is not lost, the false positive rate is low, the binding capacity of the screened aptamer is high, the library preparation time is short, only one round of enrichment can be performed, the library preparation repeatability is high, and the method is suitable for an automatic mechanical arm are achieved.

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.