42 results about "Template switching" patented technology

The present invention combines the use of template switching with nucleic acid amplification—such as PCR—to generate amplification products representative of an entire RNA population. An RNA polymerase promoter sequence allows transcription-based amplification to be performed on the derived amplification products such that antisense amplified RNA (aRNA) or complementary RNA (cRNA) is produced for subsequent downstream applications.

The invention provides a device for intelligently configuring an operation interface of electronic equipment, which is connected with a server through a transmission unit arranged in the device. The device comprises a central processing unit, an interface template unit and a template switching unit, wherein the interface template unit and the template switching unit are respectively connected with the central processing unit; the interface template unit is used for storing a plurality of operation interface templates scheduled by the central processing unit, and each operation interface template comprises a plurality of application programs; the template switching unit is used for sending a switching signal to the central processing unit so as to schedule a corresponding interface template; a clock unit is used for supplying a user use operation interface template and the time of a corresponding application program to the central processing unit; and the central processing unit is used for accumulatively recording use habits of the electronic equipment at different times and different places and use habits of the application program on the operation interface and intelligently regulating the operation interface of the electronic equipment according to the use habits to ensure that the operation interface of the electronic equipment better accords with the actual requirement on a user, therefore, the user experience is effectively enhanced.

A method of preparing a DNA copy of a target polynucleotide using template switching is described. The method includes mixing a double stranded template/primer substrate made up of a DNA primer oligonucleotide associated with a complementary oligonucleotide template strand with a target polynucleotide in a reaction medium and adding a suitable amount of a non-retroviral reverse transcriptase to the reaction medium to extend the DNA primer oligonucleotide from its 3′ end to provide a DNA copy polynucleotide. The DNA copy polynucleotide includes a complementary target DNA polynucleotide that is synthesized using the target polynucleotide as a template. Methods of adding nucleotides to the double stranded template/primer substrate are also described. The method can be used to facilitate detection, PCR amplification, cloning, and determination of RNA and DNA sequences.

Disclosed herein include systems, methods, compositions, and kits for 5′-based gene expression profiling. Some embodiments provide synthetic particles (e.g., beads) associated with a first plurality of oligonucleotide barcodes and a second plurality of oligonucleotide barcodes. In some embodiments, nucleic acid targets (e.g., mRNAs) are initially barcoded on the 3′ end with the first plurality of oligonucleotide barcodes and subsequently barcoded on the 5′ end following a template switching reaction and intermolecular hybridization with the first plurality of oligonucleotide barcodes and extension. Immune repertoire profiling methods are also provided in some embodiments.

The present invention provides a new approach to creating novel polynucleotide sequences by point mutation and recombination in vitro of a set of parental sequences. The new polynucleotide sequences can be useful in themselves or they can be used for the templated synthesis of polymers, e.g. polypeptides composed of α-amino acids as occurring naturally in protein synthesis or polymers comprised of β-amino acids or other building blocks as described (Templated molecules).

Disclosed herein include systems, methods, compositions, and kits for labeling nucleic acid targets in a sample. In some embodiments, nucleic acid targets (e.g., mRNAs) are initially barcoded on the 3′ end and are subsequently barcoded on the 5′ end following a template switching reaction and intermolecular and/or intramolecular hybridization and extension. 5′- and/or 3′-barcoded nucleic acid targets can serve as templates for amplification reactions and/or random priming and extension reactions to generate a sequencing library. The method can comprise generating a full-length sequence of the nucleic acid target by aligning a plurality of sequencing reads. Immune repertoire profiling methods are also provided in some embodiments.

The invention discloses a method for achieving user-defined graphical user interface. The method includes setting a template pack catalog in a graphical user interface material catalog; respectively storing templates in a folder manner according to template names in the template pack catalog, setting a configuration file of the user-defined graphical user interface in the graphical user interface material catalog, and setting template option values in the configuration file; adding an image file designating naming rules in a designation material catalog, wherein the image file is identified as an icon of the designation material catalog or file; reading the template option values in the configuration file, loading materials of each optional template, displaying preview images and template switching buttons of each optional template, and selecting corresponding templates according to operations of a user; reading the configuration file and catalog and file listings in the graphical user interface material catalog, identifying configuration items and the naming rules, correspondingly processing each rule and configuration, and displaying the graphical user interface according to processing results. The invention further discloses a device for achieving the user-defined graphical user interface.

This application discloses methods for cDN′A synthesis with improved reverse transcription, template switching and preamplification to increase both yield and average length of cDNA libraries generated from individual cells. The new methods include exchanging a single nucleoside residue for a locked nucleic acid (INA) at the TSO 3′ end, using a methyl group donor, and/or a MgCb concentration higher than conventionally used. Single-cell transcriptome analyses incorporating these differences have full-length coverage, improved sensitivity and accuracy, have less bias and are more amendable to cost-effective automation. The invention also provides cDNA molecules comprising a locked nucleic acid at the 3′-end, compositions and cDNA libraries comprising these cDNA molecules, and methods for single-cell transcriptome profiling.

The invention relates to a template switching method for construction of an integrated process of product reliability, maintainability and supportability (RMS) and performance. The template switching method comprises the following steps: (1) selecting an integrated process template according to the product, and by adding, deleting and modifying, constructing an initial integrated process to be optimized; (2) by using the bilateral switching relation between a process directed graph and a design structure matrix, constructing an initial design structure matrix N on the basis of determining the information dependency relation among working items; (3) solving an adjacent matrix A and a reachable matrix R corresponding to N, and a product of R and a transposed matrix RT thereof to find out a coupled set in the matrix; (4) sequencing the working items in the coupled set by a segmentation algorithm, an aggregation algorithm and a tearing algorithm; (5) regarding the coupled set as a working item to be sequenced and graded with other independent items, and forming the design structure matrix after optimization; (6) by using the bilateral switching relation between the process directed graph and the design structure matrix, switching the design structure matrix after optimization to be the integrated design process of RMS and performance to be determined; and (7) inspecting the integrated process of RMS and performance to be determined by a standard inspection list, and forming a new integrated process template for later use.

The invention discloses a three-generation library constructing and sequencing method for overall length amplification of a BCR immunity group library. The method comprises the steps of (1) designingand synthesizing primers according to the constant region of a BCR consistency sequence; (2) synthesizing a first chain cDNA under the action of an Oligo dT primer and a BCR Template Switching Oligo primer; (3) performing first-round amplification and second-round amplification on the overall length of BCR cDNA; (4) mixing samples of BCR overall length amplicon fragments; (5) performing library construction; and (6) performing three-generation sequencing. According to the method disclosed by the invention, multiple primers are designed in the conservative region of a BCR constant region, so that the overall length sequences of the whole BCR are covered. During BCR sequence amplification, in order to further improve the validity of 3' end amplification, multiple groups of semi-nest primersare designed, and through matching and screening of the semi-nest primers, amplicon fragments covering the whole BCR overall length can be obtained. Through a PacBio library constructing technologicalprocess, sequencing contacts having barcode are added to the two ends of each amplicon, and then PacBio sequencing can be performed; and through splitting and data correction, HiFi Reads of which theaccuracy rate is 99% or above can be obtained.

Disclosed herein, are compositions, methods, and kits comprising engineered reverse transcription enzymes that exhibit several desired properties such as thermal stability, processive reverse transcription, non-templated base addition, and template switching ability. The engineered reverse transcription enzymes described herein demonstrate unexpectedly higher resistance to cell lysate inhibition,greater ability to capture full-length mRNA transcripts, and demonstrate improved results in small reaction volumes as compared to other engineered reverse transcription enzymes.

The invention discloses a high-throughput sequencing library construction method for trace free RNA of body fluid. The method comprises the following steps: purifying a body fluid sample exRNA with asilicon substrate, performing dephosphorylation, adding polyA tails, and performing inverse transcription on tailing products by using template switching and nucleic acid locking technologies so as toobtain cDNA of which the 5'-end is provided with linkers; and performing low-cycle PCR (Polymerase Chain Reaction), and finally obtaining the high-throughput sequencing library. According to the method disclosed by the invention, by using the template switching and nucleic acid locking technologies, deep optimization is performed on problems encountered by trace RNA in the process of constructingthe high-throughput sequencing library, and the library construction time and cost can be greatly reduced. By using the technology in the invention, library construction can be completed on less than10ng of trace RNA in one day, high-quality data can be acquired by virtue of high-throughput sequencing, and many existing disease-related RNA biomarkers can be found by analysis.

The invention discloses a vehicle body large part positioning tool with a template switching function, and belongs to the field of auxiliary positioning tools of railway vehicle body large parts. The positioning tool comprises double guide rails and a template positioning base, wherein the double guide rails consist of two identical guide rails; chutes with outer side stops are formed in the upper end surfaces of the guide rails; the two identical guide rails are symmetrically arranged in parallel; the template positioning base comprises two square baffle plates and a clamping plate component; the clamping plate component comprises four clamping plates with through grooves, and multiple template positioning screw holes; and the four clamping plates are integrally molded, and are arranged at intervals by circumference angles. Vehicle body large part templates can quickly finish the switching of selected template stations according to actual requirements, so that the operation of replacing the vehicle body large part templates in batches is convenient and quick, the production efficiency can be largely improved, the production cost is saved, and the benefit is created. In addition, the positioning tool has such advantages as simple and practical structure, convenience for operation, low cost and convenience for promotion and popularization.

High throughput methods and compositions (e.g., kits) for the amplification of RNA fragments, including in particular, for the detection of fusion mutations in a high volume of samples, e.g., by high throughput sequencing method. These methods may include barcoding cDNA preparations with template switching reactions, indexing pools of libraries and intensive use of automatic liquid handling, and providing a ready-to-sequence library mix.