911 results about "Polymerase" patented technology

Disclosed herein are certain thiadiazolyl derivatives Formula (I):that inhibit DNA Polymerase Theta (Polθ) activity, in particular inhibit Polθ activity by inhibiting ATP dependent helicase domain activity of Polθ. Also, disclosed are pharmaceutical compositions comprising such compounds and methods of treating and / or preventing diseases treatable by inhibition of Polθ such as cancer, including homologous recombination (HR) deficient cancers.

The invention discloses an oil palm U6 promoter gene and application thereof, and belongs to the technical field of biology. The nucleotide sequence of the promoter gene is shown as SEQ ID NO.1. The oil palm RNA polymerase III type promoter gene, namely the oil palm endogenous U6 promoter gene EgU6, is obtained by cloning in an oil palm genome for the first time, and the promoter gene has high transcriptional activity and can drive downstream fluorescent protein mNeonGreen expression. The candidate oil palm endogenous U6 promoter gene can be provided for subsequently establishing a high-efficiency oil palm gene editing technology system based on a CRISPR / cas9 (Clustered Regularly Interspaced Short Palindromic Repeats / Cas9) system.

The invention discloses a kit for detecting miRNAs of multiple extracellular vesicles based on a TtAgo-driven rolling circle amplification technology and application of the kit, a platform combines TtAgo protein and RCA reaction, cooperates with endonuclease activity and DNA polymerase activity of TtAgo, accurately controls reaction kinetic balance between temperature and enzyme activity, not only does not damage the cutting activity of TtAgo, but also can detect miRNAs of multiple extracellular vesicles. The method provided by the invention can be used for detecting multiple extracellular vesicles miRNAs, ensures strong polymerization amplification performance, creates a strong one-step single-tube experiment, greatly simplifies the experimental process, shortens the detection time, realizes ultra-sensitive and specific detection of nucleic acid markers, and can be used for detecting multiple extracellular vesicles miRNAs.

The invention provides an RNA polymerase variant, the enzyme specific activity of the variant is obviously improved compared with that of a wild type, and the addition amount of polymerase is reduced, so that the cost is saved. In addition, the invention further provides a method for generating RNA through in-vitro transcription.

The present disclosure relates to Taq DNA Polymerase mutants comprising at least three mutations which are capable of high levels of DNA amplification in the presence of several PCR inhibitors and compositions comprising the same. Also disclosed are methods of performing mutagenesis on wild-type Taq DNA polymerase to obtain the disclosed Taq DNA polymerase mutants, DNA amplification using the same, and cDNA amplification using the same.

The invention provides an RNA polymerase variant and application thereof, and relates to the technical field of biology. The RNA polymerase variant provided by the invention has the performance of remarkably improving the integrity of a super-long fragment mRNA transcription product, can be used for preparing mRNA through in-vitro transcription, particularly can improve the integrity of the product when preparing saRNA exceeding 10000 nt, and has important significance for industrial production of saRNA.

The present document describes methods and systems for amplifying and quantifying amplification of a nucleic acid molecule, with a polymerase chain reaction (PCR) or a loop-mediated isothermal amplification (LAMP), by irradiating, with a heating activation light beam from a continuous wave laser a biological enzymatic reaction mixture in solution comprising a nucleic acid template, a polymerase enzyme, and chemically modified nanoparticles. Quantification of the amplification is achieved by irradiating the biological enzymatic reaction mixture during an annealing and / or elongation steps with an ultraviolet (UV) light source and measuring with a photodetector a transmission change in UV light transmission.

The invention discloses a Taq DNA polymerase mutant with improved excision activity and application thereof, and belongs to the technical field of enzyme engineering. The invention aims to solve the technical problems of low probe cutting efficiency, weak fluorescence signal, limited detection sensitivity (especially for low-abundance targets), low reaction speed (high Ct value) and reduced specificity caused by insufficient 5 '-> 3' excision enzyme activity of wild Taq DNA polymerase in TaqMan probe method qPCR (quantitative polymerase chain reaction). According to the technical scheme, the Taq DNA polymerase mutant with the high 5 '-3' exonuclease activity is provided, the amino acid sequence of the Taq DNA polymerase mutant is shown as SEQ ID NO.2, the exonuclease activity of the mutant is improved by 77%, the cutting speed of a TaqMan probe is greatly increased, accumulation of fluorescence signals is accelerated, the Ct value is reduced, and the detection time is remarkably shortened.

The invention discloses a variable region amino acid sequence of a monoclonal antibody of a West Nile virus non-structural protein NS1 and application of the variable region amino acid sequence, and belongs to the technical field of medicines. According to the invention, West Nile virus non-structural protein NS1 expressed by human embryo kidney 293 cells is used as an antigen to immunize a rabbit, B cells capable of being specifically combined with the West Nile virus non-structural protein NS1 are screened from rabbit spleen cells through flow sorting, and a signal peptide and a variable region gene fragment of an antibody are cloned through reverse transcription-polymerase chain reaction; according to the present invention, the non-structural protein NS1 of flaviviridae flaviviridae virus is taken as a template, and is connected with a constant region gene to an expression vector, and after mammalian cell expression and purification, the monoclonal antibody which has high affinity and is not combined with the non-structural protein NS1 of other eight viruses of flaviviridae flaviviridae virus is obtained through enzyme-linked immunosorbent assay; the monoclonal antibody has application value in diagnosis and prevention and treatment of West Nile virus infection.

The invention relates to the technical field of biology, in particular to a cis-trans typing method for drug-resistant gene mutation EGFR T790M / C797S. The invention provides an EGFR (Epidermal Growth Factor Receptor) T790M / C797S cis-trans typing method based on a locked nucleic acid LNA (Low Nucleic Acid) probe, ultrahigh-specificity DNA (Deoxyribose Nucleic Acid) polymerase and a primer probe combination. The typing method can be used for accurately distinguishing EGFR T790M / C797S double-mutation cis-configuration and trans-configuration; meanwhile, the kit has the advantages of high sensitivity, high specificity, simple steps and low cost, is suitable for detecting low-abundance samples such as circulating tumor DNA (ctDNA) and can meet the clinical rapid detection requirement.

The invention belongs to the field of nucleic acid detection and molecular biology, and particularly relates to a method and a kit for eliminating false positive results in nucleic acid amplification reaction. According to the method, after a nucleic acid amplification reaction (taking recombinase polymerase amplification, namely RPA, for example) is completed, a target amplification product is subjected to selective enzyme digestion by utilizing restriction endonuclease, non-target products (such as primer dimers and non-specific amplification products) are not cut, and meanwhile, the non-target products which are not subjected to enzyme digestion are removed by combining solid-phase separation, so that the target amplification product is obtained. Therefore, the false positive result is eliminated. The invention also discloses a kit containing the restriction enzyme. According to the method, the specificity and the signal-to-noise ratio of a nucleic acid amplification reaction, especially RPA, are remarkably improved, the cost is low, operation is easy, the method is compatible with an existing technical platform, the method is suitable for detection scenes such as clinical molecular diagnosis, environmental monitoring and food safety, and the problem of misjudgment caused by false positive signals is effectively avoided.

The invention discloses an anti-Taq DNA polymerase antibody or an antigen binding fragment thereof and application thereof, and relates to the technical field of biology. The antibody or the antigen binding fragment of the antibody can be specifically bound with Taq DNA polymerase, the polymerization activity and / or excision activity of the Taq DNA polymerase are / is blocked, and the stability of the Taq DNA polymerase is improved.

The present invention provides a process for the production of a closed linear DNA comprising the steps of (a) providing a DNA template comprising a DNA sequence of interest; (b) amplifying DNA from the DNA template of step (a) wherein the amplification is primed with a primase / polymerase enzyme; (c) generating a closed linear DNA with the amplified DNA produced in step (b); and (d) purifying the closed linear DNA produced in step (c). The invention also provides a closed linear DNA obtainable according to the process of the invention, a pharmaceutical composition comprising a therapeutically effective amount of the closed linear DNA of the invention, and a concatameric DNA comprising repeats of a DNA sequence of interest.

The invention provides an RNA (Ribonucleic Acid) polymerase variant and a preparation method thereof, and an RNA product with low dsRNA pollution can be obtained by applying the RNA polymerase variant disclosed by the invention to in-vitro transcription. In addition, the invention further provides a method for preparing RNA through in-vitro transcription.

The invention belongs to the technical field of biology, and relates to an RNA polymerase variant, and when the RNA polymerase variant is applied to in-vitro transcription, an RNA product with low dsRNA pollution can be obtained. In addition, the invention further provides a method for generating RNA through in-vitro transcription.

The application discloses a composition, a kit, a use and a method for removing nucleic acid residues in enzyme products. The application has the advantages that, compared with the prior art, the application can effectively remove host cell nucleic acid contamination in a polymerase, and particularly, through a specific sgRNA design of a target sequence, it is ensured that even if a residual part of the sequence does not affect subsequent microbial detection. In addition, the method is simple to operate, has high purification efficiency, and is suitable for large-scale polymerase production and application.

The embodiment of the invention discloses a preparation method of a kit applied to endometrial cancer microsatellite state detection. The preparation method comprises the following steps: taking a first preset volume of polymerase chain reaction amplification liquid; taking a polymerase chain reaction primer mixed solution with a second preset volume; and combining the weighed polymerase chain reaction amplification liquid and polymerase chain reaction primer mixed liquid into the kit applied to endometrial cancer microsatellite state detection. According to the embodiment, the accuracy and the stability of microsatellite state detection of the endometrial cancer can be improved, so that the kit consumed by repeatedly detecting the microsatellite state in the endometrial cancer is reduced.

The invention provides a polypeptide for activating cellular immunity in chronic hepatitis B and application of the polypeptide, and belongs to the technical field of biological medicine. A polypeptide library is designed and synthesized on the basis of a preS1 structural domain for coding HBV large HBsAg, a full-length core protein Core, a polymerase protein fragment rich in T cell epitopes and an mRNA-PreS1CPX holoantigen sequence (as shown in SEQ ID NO.1) of full-length X protein HBX, and peptide fragments capable of activating T cell immunity are screened by utilizing ELISPOT and flow cytometry. Experimental results show that the polypeptide sequences as shown in SEQ ID NO.2-15 can promote HBV antigen specific immune response by stimulating CD8 + T lymphocytes to secrete IFN-gamma, so that immune activation treatment of hepatitis B is realized.

The application discloses a kind of compositions, reagents and its detection method for high-throughput gene sequencing, belong to gene sequencing technical field, the composition includes Tris-HCl buffer 80-120 parts, potassium chloride 40-60 parts, ammonium sulfate 10-20 parts, magnesium chloride 15-25 parts, dNTPs mixed solution 30-50 parts, heat-stable DNA polymerase mutant 8-15 parts, PCR promoter complex 5-12 parts, modified iridium quinoline ketone complex 1-5 parts, glycerol 150-250 parts, Tween-20 1-3 parts, EDTA disodium salt 0.5-2 parts and nuclease-free ultrapure water, by introducing the modified iridium quinoline ketone complex of innovation, effectively solve the technical problems of low amplification efficiency, poor multiplex PCR reaction uniformity and high non-specific amplification background for high GC content, complex secondary structure genomic template, significantly improve the specificity of amplification, uniformity and the quality and reliability of final sequencing data.

A method for promoting erythrocyte differentiation is provided. A method for increasing RNA polymerase II arrest mediated by the MED26 polypeptide is also provided. Furthermore, a method for delivering reagents to cells is also provided.

Disclosed herein, inter alia, are mutant enzymes, kits, and methods of use thereof.

The PCR plate according to the present invention includes: a body portion having one or more reaction wells; an insertion portion extending from the body portion, inserted into a nucleic acid extraction cartridge, and having an injection port into which a nucleic acid solution is injected; a flow path portion allowing the nucleic acid solution to flow from the injection port to the reaction wells; and a blocking portion attached to the body portion for blocking backflow of the nucleic acid solution from the reaction wells to the flow path portion side.

The invention belongs to the field of detection, and particularly relates to a kit for detecting and identifying staphylococcus capitis and a detection method. The kit for detecting and identifying the staphylococcus capitis comprises Taq DNA (deoxyribonucleic acid) polymerase, a 10 * Taq buffer solution, a dNTP (deoxyribonucleoside triphosphate) mixed solution, an upstream primer, a downstream primer and a DNA Marker III. During detection, DNA of a sample to be detected is taken as a template, electrophoresis is performed after PCR amplification, if a 863bp band appears, the sample is positive, rapid and specific detection can be performed, and clinical (such as non-lactation period mastitis) requirements are met.

The present disclosure provides variant RNA polymerases with amino acid substitutions, and their use of which increases transcription yield, RNA integrity, capping efficiency, and reduces dsRNA formation during an in vitro transcription reaction.

The invention provides a ready-to-use PCR tube preloaded with Taq polymerase and reaction components and stable at room temperature, and belongs to the field of molecular biology and nucleic acid diagnosis. The invention provides a ready-to-use PCR (polymerase chain reaction) tube which can be stored at room temperature and is pre-filled with a premix for carrying out polymerase chain reaction (PCR) amplification on all necessary components in advance. The kit is characterized in that each tube is pre-filled with locally prepared Taq polymerase, a PCR buffer solution, dNTPs, MgCl and target specific primers (only a customized PCR kit-ready-to-use PCR plus tube contains the primers), and all the components are dried and stored in a dry powder form, so that the shelf life can be prolonged, and the tedious steps of on-ice operation can be avoided. According to the method, the error rate in the PCR preparation process is reduced to the maximum extent, and the diagnosis and research work flow is simplified.

A fiber optic device and method for uniform illumination of biological samples distributed over one or more microcell arrays of a microwell plate can establish more reliable polymerase chain reaction (PCR) data sampling. In some embodiments, the apparatus includes a light engine that emits a light beam captured by a fiber optic bundle for delivering one or more light beams to each microchamber; each light beam thereby fluoresces the biological sample. The fiber optic cable of the fiber optic bundle comprises a distal output end held in place using a mechanical support for flexibly aligning the output beam of the distal output end to be arranged such that the center of the output beam is directed to different locations in the one or more microcell arrays, wherein each of adjacent ones of the output beams partially overlaps so that the light distribution is uniform.

The present invention relates to cells comprising an orthogonal DNA replication machine, linear plasmids replicable by said machine, uses of said cells and said linear plasmids, methods of maintaining linear plasmids in cells, methods of evolving sequences of interest, and methods of preparing polypeptides or nucleic acids. The invention also relates to error-prone DNA polymerases, nucleic acids encoding said polymerases and uses thereof.

The present disclosure provides an application method in rice callus differentiation based on an Oryza sativa Leafy Cotyledon 1 (OsLEC1) gene, including the steps of: selecting guide ribonucleic acid (gRNA) target sites; cloning tandem fragments including gRNA; ligating each gRNA fragment; performing polymerase chain reaction (PCR) amplification on a ligation product; performing enzyme digestion on the purified product and a target vector; transforming the ligated vector; performing Agrobacterium-mediated genetic transformation of rice; and screening and identifying transgenetic plants. In the present disclosure, the differentiation of callus directly affects the emergence efficiency of transgenic plants. The knockout of OsLEC1 can promote the differentiation of rice callus, suggesting that OsLEC1 can serve as an important target gene for improving the transformation efficiency of rice and even gramineous crops. OsLEC1 can be used as a starting point to construct various molecular tools to enhance transformation efficiency.