22 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.

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.

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.

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 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.

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.

This disclosure provides methods and compositions relating to the characterization of the chikungunya virus genome, including specific sets of oligonucleotide primers for the amplification of the chikungunya virus genome for downstream sequencing, alignment, and identification of variants in the chikungunya virus genome.

The present application relates to a single fluorescence method and a double fluorescence method for rapid detection of Cronobacter dublinensis and Cronobacter univans by ERA technology. The present application also relates to an oligonucleotide primer probe composition for the method. The present application also relates to a detection kit comprising the composition. The ERA fluorescence method detection using the composition of the present application can simply, rapidly, specifically and sensitively detect Cronobacter dublinensis and Cronobacter univans.

The present application relates to the single fluorescence method and double fluorescence method for rapid detection of Cronobacter sakazakii and Cronobacter zurichensis by using ERA technology. The present application also relates to the oligonucleotide primer probe composition for the method. The present application also relates to the detection kit comprising the composition. The ERA fluorescence method detection using the composition of the present application can simply, rapidly, specifically and sensitively detect Cronobacter sakazakii and Cronobacter zurichensis.