40 results about "Oligonucleotide Primer" patented technology

The invention provides a system and methods of multiplexed, solid-phase isothermal nucleic acid amplification. In various aspects, the invention uses a microfluidic device that includes a field of actuatable microposts in a reaction (or assay) chamber to enhance fluid flow, mixing, and hybridization / capture efficiency in a solid-phase capture assay. In various other aspects, the invention uses oligonucleotide primers immobilized in a field of actuatable microposts in a reaction chamber of a microfluidics device for capture and amplification of target-specific nucleic acids in a sample fluid. The invention provides methods of producing a micropost field (array) on a substrate for printing of a capture array (e.g., an array of primer spots). The invention also provides methods of printing an array of capture spots (e.g., primer spots) on the substrate surface of a micropost field.

A composition for identifying a hemp cultivar Pinkpepper or a progeny plant thereof includes the oligonucleotide primer set of SEQ ID NOs: 7 and 8, and / or the oligonucleotide primer set of SEQ ID NOs: 9 and 10. A method for identifying hemp cultivar Pinkpepper or a progeny plant thereof includes extracting genomic DNA from a hemp plant, amplifying a target sequence by having the extracted genomic DNA as template and carrying out an amplification reaction using the oligonucleotide primer set, and detecting a product produced by the application of the target sequence.

Flow cells and methods are described that enable control over DNA fragment length bias during amplification. One example flow cell includes a substrate and a polymeric hydrogel positioned over at least a portion of the substrate. A plurality of oligonucleotide primers may be attached to the polymeric hydrogel, wherein the plurality of oligonucleotide primers defines a density gradient across the substrate of the flow cell.

The invention provides methods for simultaneously enriching multiple target regions of interest in one reaction volume, from cell-free DNA isolated from a blood or plasma sample, followed by high-thought sequencing and sequence read analysis. The invention also provides library of target-specific oligonucleotide primers or probes for the multiplexed target enrichment.

The present disclosure provides for methods of generating a single stranded primer from an oligonucleotide probe. For instance, the present disclosure is directed to a method of preparing oligonucleotide primers from oligonucleotide probes, such as oligonucleotide probes which have been amplified, and which include a target specific primer sequence, a forward primer binding site, and a reverse primer binding site. It is believed that the presently disclosed method permits the synthesis of thousands to hundreds of thousands and even a million primers in a cost-effective manner, with good yield, in high quantities and at high purity levels.

The present application relates to a kind of oligonucleotide primer probe compositions for detecting Pseudomonas syringae, the pathogen of vegetable leaf spot disease based on ERA fluorescence method or ERA test strip method, and the detection method using the composition.The present application can simply, quickly, specifically, sensitively and visually broad-spectrum identify a variety of pathogenic variants of Pseudomonas syringae, the pathogen of vegetable leaf spot disease, the method is easy to operate, reaction time is short, without complex temperature control equipment, can be widely applied to the on-site rapid screening and disease diagnosis of Pseudomonas syringae in vegetable sample.

Disclosed herein are novel methods and compositions for rapidly extracting and amplifying nucleic acids from a sample where the sample is combined with an extraction reagent comprising a reducing agent to form a mixture and incubating said mixture at ambient temperature for a period of time not exceeding 30 minutes to generate a nucleic acid extract. In certain embodiments of the method, the nucleic acid extract is subjected to a nucleic acid amplification reaction. In certain aspects, oligonucleotide primers specific for nucleic acids of Chlamydia species and / or Neisseria species are added prior to initiating the amplification reaction.

The embodiment of the invention discloses a TaqMan real-time fluorescent quantitative PCR probe primer combination for identifying tilapia mossambica, a kit and application. Comprising a probe, an upstream primer and a downstream primer, the nucleotide sequence of the probe is 5 '-FAM-CGCTATACAGGG-MGB-3', the nucleotide sequence of the upstream primer is 5 '-GACTTTCCCCCATATTC-3', and the nucleotide sequence of the downstream primer is 5 '-GGGGTAGACAGTTCACCCTGTA-3'. Based on difference of mitochondrial control areas of different fishes, the TaqMan probe real-time fluorescent PCR detection method established by designing specific oligonucleotide primers and probes can effectively distinguish tilapia homosambica from other fishes, has excellent specificity and sensitivity in the aspect of identifying tilapia homosambica species, and can be used for detecting the species of tilapia homosambica. The detection accuracy is greatly improved, meanwhile, the detection time is shortened, the effects of stability, reliability, short period and low cost are achieved, and the actual application requirements are met.

Provided herein are methods and compositions for the synthesis of 5 '-capped RNA wherein the starting capped oligonucleotide primer has the general form m7Gppp [N2' Ome] n [N] m wherein m7G is an N7-methylated guanosine or any guanosine analog, N is any natural, modified or non-natural nucleoside, "n" may be any integer from 0 to 4 and "m" may be an integer from 1 to 9.

The present invention relates to the detection of modifications in polynucleotides by binding an affinity binding element to a nucleotide sequence. The affinity binding element may include a compound that binds to a modified nucleotide base and a nucleotide sequence complementary to a target nucleotide sequence, an oligonucleotide primer complementary to the target nucleotide sequence, or a nucleotide sequence complementary to an oligonucleotide linker. The present invention provides methods for detecting one or more modifications in a nucleotide sequence. The invention also provides affinity binding elements, and optionally, oligonucleotide primers and / or oligonucleotide probes, and methods of using the affinity binding elements in assays to detect modified bases. The methods of the invention enable the detection of modified nucleotides associated with cancer, psychiatric disorders or metabolic diseases.

The present invention is directed to a method for detecting the presence or absence of monkeypox virus (MPXV) in a biological sample, the method comprising: a) contacting the sample or nucleic acid extracted from said sample with reagents and an oligonucleotide combination capable of amplifying a target region of a monkeypox viral genome, wherein said oligonucleotide combination is selected from the group consisting of: i) a B7R oligonucleotide primer set specific for amplifying a target sequence in the B7R gene of the MPXV genome generating an MPXV amplification product comprising the sequence of SEQ ID NO:65; and ii) a G2R oligonucleotide primer set specific for amplifying a target sequence in the G2R gene of the MPXV genome generating an MPXV amplification product comprising the sequence of SEQ ID NO:63; b) performing an in vitro nucleic acid amplification reaction, wherein any MPXV nucleic acid present in the sample is used as a template to generate the MPXV amplification product corresponding to the oligonucleotide combination of i) or ii); and c) detecting the presence or absence of the MPXV amplification product generated in step b), wherein the presence of the MPXV amplification product is indicative of the presence of MPXV in the biological sample. The present invention also provides oligonucleotides which can be used in the method of the invention as primers and probes.

The present disclosure relates generally to devices and methods for rapid and accurate diagnosis of single nucleotide polymorphisms. According to one or more embodiments, a system for identifying a single nucleotide polymorphism (SNP) comprises an oligonucleotide primer having a SNP recognition sequence, a fluorophore conjugated to the oligonucleotide primer at a conjugation site to form a fluorophore-conjugated oligonucleotide primer, a polymerase for extending the oligonucleotide primer to form a double-stranded deoxyribonucleic acid (DNA) amplicon upon binding to an oligonucleotide strand having a complementary sequence to the oligonucleotide primer, and wherein the fluorophore-conjugated oligonucleotide primer is the only primer in the system such that the fluorophore emits a fluorescent signal only when the fluorophore-conjugated oligonucleotide primer binds to a sequence specifically complementary to the SNP recognition sequence.

The invention relates to a set of oligonucleotide primers, and kits and in vitro diagnostic devices comprising the set of oligonucleotide primers. The set of oligonucleotide primers, kits and in vitro diagnostic devices can be used in detecting Neisseria gonorrhoeae nucleic acid and diagnosing Neisseria gonorrhoeae infection with high sensitivity and specificity. The invention also relates to methods of loop-mediated isothermal amplification, methods of detecting Neisseria gonorrhoeae nucleic acid, and methods of diagnosing Neisseria gonorrhoeae infection using the set of oligonucleotide primers or the kits.

Disclosed are oligonucleotide primers that hybridize specifically with any base sequence designed from the base sequences of the N gene, RNA-dependent RNA polymerase gene, M gene, and S gene of SARS-COV-2, a nucleic acid amplification method using said primers, a test method for SARS-COV-2 infection by detection of nucleic acid amplification, and a COVID-19 test kit.

The present invention relates to methods for the detection of nucleic acids of defined sequence and kits and devices for use in said methods. The methods employ restriction enzymes. polymerase and oligonucleotide primers to produce an amplification product in the presence of a target nucleic acid. which is contacted with oligonucleotide probes to produce a detector product.

The present invention provides methods and compositions for determining the presence and / or amount of pathogenic Leptospira in a test sample. In particular, substantially purified oligonucleotide primers and probes are described that can be used for qualitatively and quantitatively detecting pathogenic Leptospira nucleic acid in a test sample by amplification methods. The present invention also provides primers and probes for generating and detecting control nucleic acid sequences that provide a convenient method for assessing internal quality control of the Leptospira assay.

This invention claims processes that append a single nucleotide having a 3′-ONH2 moiety to the 3′-ends of an oligonucleotide primer using 3′-deoxynucleoside triphosphates that have, instead of a 3′-OH moiety, a 3′-ONH2 moiety, where the nucleotides contain both standard and non-standard nucleobases, and where as a key claim limitation, substantially no hydroxylamine is present in the solutions used in the claimed processes.

Provided herein are methods and compositions for synthesizing 5'Capped RNAs wherein the initiating capped oligonucleotide primers have the general form m7Gppp[N2'Ome]n[N]m wherein m7G is N7-methylated guanosine or any guanosine analog, N is any natural, modified or unnatural nucleoside, "n" can be any integer from 0 to 4 and "m" can be an integer from 1 to 9.

The invention relates to a set of oligonucleotide primers, and kits and in vitro diagnostic devices comprising the set of oligonucleotide primers. The set of oligonucleotide primers, kits and in vitro diagnostic devices can be used in detecting Chlamydia trachomatis nucleic acid and diagnosing Chlamydia trachomatis infection with high sensitivity and specificity. The invention also relates to methods of loop-mediated isothermal amplification, methods of detecting Chlamydia trachomatis nucleic acid, and methods of diagnosing Chlamydia trachomatis infection using the set of oligonucleotide primers or the kits.

A method of amplifying a target nucleic acid in a DNA sample comprising (a) contacting the DNA sample containing the target nucleic acid with a DNA polymerase, at least two oligonucleotide primers designed to flank the target nucleic acid, a mixture of dATP, dGTP, dCTP, and dTTP, and a conjugate comprising a DNA polymerase inhibitor covalently attached to a negative temperature sensitive polymer, (b) heating the output of step (a) to a temperature at which the conjugate precipitates and thus the DNA polymerase is no longer inhibited and (c) amplifying the target nucleic acid, e.g. by performing PCR steps of denaturing the target nucleic acid, annealing the primers to the target nucleic acid, and extending the primers, wherein step (c) is repeated at least two times. Also provided is a conjugate comprising a DNA polymerase inhibitor covalently attached to a negative temperature sensitive polymer and kits and aqueous compositions comprising the same.

The present disclosure relates, in general, to the methods for the rapid detection of the presence or absence of Lymphogranuloma Venereum (LGV)-causing serovars of Chlamydia trachomatis in a biological or non-biological sample. The methods can include performing an amplification step, a hybridization step, and a detection step. Furthermore, oligonucleotide primers and probes targeting the pmpH gene for the L serovars of Chlamydia trachomatis, along with kits are provided that are designed for the detection of L serovars.

Methods for strand-displacement amplification of a target nucleic acid have been developed. Compositions for use according to the methods are also provided. The methods typically include steps of removing, hybridizing and extending a selectively removable or digestible primer with a strand-displacing DNA polymerase once or more than once to provide a multiplicity of copies of the target nucleic acid. Methods for strand-displacement amplification of single or double-stranded target DNA molecules are provided. In some forms, when the methods include amplification of a double-stranded nucleic acid, the methods employ one or more adapter handles that perform the function of providing a removable region on a single strand of the ds target. Compositions and kits of oligonucleotide primers and double-stranded nucleic acid adapters including selectively removable regions are also provided for use in the described methods.

The invention relates to a fluorescence detection method for respectively and rapidly detecting Cronobacter mojinensis and Cronobacter kangdi by adopting a body temperature amplification (ERA) technology. The invention also relates to an oligonucleotide primer probe composition for the method. The invention also relates to a detection kit containing the composition. When the composition provided by the invention is used for ERA fluorescence method detection, cronobacter mojinensis and cronobacter kangdi can be simply, quickly, specifically and sensitively detected.

Compositions for the detection of contaminant enzymes are provided. In particular, oligonucleotide primers are provided, wherein the primer is conjugated to a blocking moiety at its 3'end, wherein the blocking moiety comprises a cleavage site for an enzyme, and wherein cleavage at the cleavage site releases the blocking moiety and produces a 3' hydroxyl group on the oligonucleotide primer. Also provided is an oligonucleotide primer comprising a cleavable moiety at its 5' end, wherein the cleavable moiety is also conjugated to an oligonucleotide tail. Also provided are methods for making the disclosed oligonucleotide primers and methods for using the oligonucleotide primers to detect a contaminant enzyme in a sample.

Provided herein are methods and compositions relating to the characterization of the dengue virus genome. The disclosure provides oligonucleotides primers for the amplification of the dengue virus genome for downstream sequencing, alignment, and identification of variants in the dengue virus genome.

The invention belongs to the technical field of gene engineering, and relates to a method for improving the expression efficiency of penicillin G acylase protein by modifying a signal peptide. According to the invention, E.coli BL21 (DE3) / pET28a-kcPGA is taken as an original strain, an Erwinia carotovora PelB amino terminal leader sequence (the signal peptide is named as pelB) is synthesized through oligonucleotide primer fusion PCR, then the pelB signal peptide is connected with a PGA gene segment without an original signal peptide by using a homologous recombination method, and finally a recombinant plasmid is constructed through seamless cloning, so that the Erwinia carotovora PelB gene fragment is obtained. And the penicillin G is transformed into escherichia coli cells, so that the expression efficiency of the penicillin G acylase is improved. Compared with original recombinant bacteria, the recombinant engineering strain constructed by the invention has the advantages that the protein expression quantity is greatly improved, the enzyme activity is also improved by about two times, the problem of low expression quantity of penicillin G acylase in escherichia coli cells is effectively solved, and the recombinant engineering strain has wider industrial production and application prospects.

Flow cells and methods are described that enable control over DNA fragment length bias during amplification. One example flow cell includes a substrate and a polymeric hydrogel positioned over at least a portion of the substrate. A plurality of oligonucleotide primers may be attached to the polymeric hydrogel, wherein the plurality of oligonucleotide primers defines a density gradient across the substrate of the flow cell.

The present disclosure provides methods and compositions of modified oligonucleotide primer and probe combinations, structurally modified with locked nucleic acids, quenchers, and dyes, effective to detect tumor-derived Human Papilloma Virus (HPV) and tumor-derived Epstein-Barr virus (EBV) and, especially, to distinguish viral DNA derived from tumors from viral DNA derived from infectious viral particles.