1247 results about "Multiplex" patented technology

The invention provides methods for simultaneously amplifying multiple nucleic acid regions of interest in one reaction volume as well as methods for selecting a library of primers for use in such amplification methods. The invention also provides library of primers with desirable characteristics, such as minimal formation of amplified primer dimers or other non-target amplicons.

Aspects of the invention relates to methods and compositions that are useful to reduce bias and increase the reproducibility of multiplex analysis of genetic loci. In some configurations, predetermined preparative steps and/or nucleic acid sequence analysis techniques are used in multiplex analyses for a plurality of genetic loci in a plurality of samples.

This invention pertains to methods, mixtures, kits and/or compositions for the determination of analytes by mass analysis using unique labeling reagents or sets of unique labeling reagents. The labeling reagents can be isomeric or isobaric and can be used to produce mixtures suitable for multiplex analysis of the labeled analytes.

The present invention generally relates to the detection of analytes, particularly biomolecules in samples. The invention also relates to compositions, methods, and kits for detecting the presence of analytes, typically in multiplex detection formats. The invention also relates to methods for determining the presence of at least one analyte in a sample, the methods employing employ single molecule detection techniques to individually detect at least one molecular complex or at least part of a molecular complex.

The present invention provides miniaturized instruments for conducting chemical reactions where control of the reaction temperature is desired or required. Specifically, this invention provides chips and optical systems for performing and monitoring temperature-dependent chemical reactions. The apparatus and methods embodied in the present invention are particularly useful for high-throughput and low-cost amplification of nucleic acids.

The invention discloses a construction method of a ctDNA ultra-low-frequency mutation detection library, a kit and an analysis method of library detection data. The construction method comprises steps as follows: S1, cfDNA is extracted from whole blood; S2, the terminal of cfDNA is restored and an A basic group is added to the 3'terminal; S3, the terminal of the cfDNA obtained in S2 is connected with connectors containing random label sequences; S4, primers for multiplex PCR (polymerase chain reaction) are designed according to the sequences of the connectors and an object region for object region capturing; S5, a PCR product in S4 is subjected to magnetic bead purification, and small-fragment DNA not subjected to non-specific amplification and primer dimer are removed; S6, a product in S5 is subjected to PCR amplification, an index sequence is introduced, and the ctDNA ultra-low-frequency mutation library is obtained. According to the method, PCR mistakes and sequencing mistakes are eliminated, and the detection specificity is improved.

The present invention provides miniaturized instruments for conducting chemical reactions where control of the reaction temperature is desired or required. Specifically, this invention provides chips and optical systems for performing and monitoring temperature-dependent chemical reactions. The apparatus and methods embodied in the present invention are particularly useful for high-throughput and low-cost amplification of nucleic acids.

The invention relates to a multiplex-PCR three-round amplification method, a primer and a kit thereof and application of the method in establishment of next-generation sequencing platform libraries. In the first two rounds of a PCR, a specific primer with the low concentration is consumed as much as possible, so that the differences of different amplicons are reduced and the homogeneity of multiplex-PCR products is improved; when the third round of the PCR is performed, adapter primers carrying different bar code sequences are added to mark different templates, the different adapter primers carry same universal amplification primers, therefore, it is guaranteed that the amplification efficiency of the different templates is consistent, and the differences of different template amplification products are reduced. Accordingly, not only is the problem that the homogeneity of ordinary multiplex-PCR amplification is poor solved, but also establishment of the sequencing libraries can be quickly and conveniently completed.

A method of in situ immunohistochemical analysis of a biological sample is provided. The method allows for the multiplex and simultaneous detection of multiple antigens, including multiple nuclear antigens, in a tissue sample.

Methods for determining the methylation status of a plurality of cytosines are disclosed. In some aspects genomic DNA target sequences containing CpGs are targeted for analysis by multiplex amplification using target specific probes that can be specifically degraded prior to amplification. The targets may be modified with bisulfite prior to amplification. In another aspect targets are cut with methylation sensitive or insensitive restriction enzymes and marked with a tag using the target specific probes. The presence or absence of methylation may be determined using methylation sensitive restriction enzyme or bisulfite treatment. Detection in many embodiments employs hybridization to tag arrays, genotyping arrays or resequencing arrays.

A system and method for developing and utilizing particle-based n-multiplexed assays that include three or more reporters utilizes n particle sets that are associated with particle identification images or labels (IDs) that differ between sets. The encoded particles for a given set are coated with a specific binding member, or in the case of the sandwich assay with coupled capture and detector binding pair members, to form particle types. The sets of particle types are then pooled, and aliquots of the particle types are removed to assay vessels. Next, samples with three or four reporter molecules are supplied to the respective vessels. After one or more incubation periods, the particles are supplied to a reader system, which determines the particle IDs to identify the particle types and also detects the reporter signals. The reader system includes multiple excitation lasers that excite the various reporters in sequence or in parallel, to supply associated signals to one or more detectors. Emission filters and wavelength discriminators are included such that a given detector receives at a given time the signals associated with a single assay binding label. The system further develops greater capacity sandwich assays by assigning subsets of capture and detector antibody pairings to the three or four reporters, respectively. The system performs greater numbers of differential RNA expressions based on the use of the three or more reporters, with one or more reporters assigned to the reference sample and the other reporters assigned to respective test samples. The system and method are also capable of performing greater numbers of SNPs utilizing primer extension reactions, by assigning different color reporters to the respective nucleotides or terminators.

Methods for balancing multiplexed PCR reactions are provided which exploit differences in primer and amplicon Tms. The methods may be controlled by a computer process. Also provided are articles of manufacture useful in such methods and compositions containing primers and probes useful in such methods.

The invention provides a method for single-tube multiplex fluorescent polymerase chain reaction (PCR) detection of human coronavirus OC43, 229E, NL63, HKU1 and SARS. The method adopts primers having sequences of SEQ ID NO: 1-10, and probes having sequences of SEQ ID NO: 11-15. The invention also provides a kit for single-tube multiplex fluorescent PCR detection of human coronavirus OC43, 229E, NL63, HKU1 and SARS. The kit contains the primers and the probes. The method provided by the invention adopts the primers which are specific primers of OC43, 229E, NL63, HKU1 and SARS, and the probes which are Taqman probes, utilizes TAMRA/CY5/FAM/ROX/JOX multiple fluorescein labeling, realizes single-tube multiplex detection of human coronavirus OC43, 229E, NL63, HKU1 and SARS, and has the advantages of strong specificity, high sensitivity, fast detection speed, simple and convenient operation and low cost. The primers and the probes can be used as detection reagents for a scientific research and clinical application.

Disclosed are compositions and a method for of amplifying nucleic acid sequences useful for detecting the presence of molecules of interest. The method is useful for detecting specific nucleic acids in a sample with high specificity and sensitivity. The method also has an inherently low level of background signal. A preferred form of the method consists of a DNA ligation operation, an amplification operation, and a detection operation. The DNA ligation operation circularizes a specially designed nucleic acid probe molecule. This operation is dependent on hybridization of the probe to a target sequence and forms circular probe molecules in proportion to the amount of target sequence present in a sample. The amplification operation is rolling circle replication of the circularized probe. A single round of amplification using rolling circle replication results in a large amplification of the circularized probe sequences. Following rolling circle replication, the amplified probe sequences are detected and quantified using any of the conventional detection systems for nucleic acids such as detection of fluorescent labels, enzyme-linked detection systems, antibody-mediated label detection, and detection of radioactive labels. Because, the amplified product is directly proportional to the amount of target sequence present in a sample, quantitative measurements reliably represent the amount of a target sequence in a sample. Major advantages of this method are that the ligation step can be manipulated to obtain allelic discrimination, the DNA replication step is isothermal, and signals are strictly quantitative because the amplification reaction is linear and is catalyzed by a highly processive enzyme. In multiplex assays, the primer oligonucleotide used for the DNA polymerase reaction can be the same for all probes. Also described are modes of the method in which additional amplification is obtained using a cascade of strand displacement reactions.

The invention discloses a multiplex amplification detection kit for detecting 60 loca of autosomes and Y-chromosomes simultaneously. The 60 loca are amplified in a multiplex manner through a polymerase chain reaction, and amplification products of the loca are detected by using a gene sequencer. The kit can be used for detecting the 60 loca of the autosomes and the Y-chromosomes simultaneously, which is a case that most STR loca can be detected by a primary reaction with a capillary electrophoresis method at present, and databases can be built for autosome STR and Y-chromosome STR simultaneously by the primary reaction. The kit has strong adaption to biomaterials, namely, one kit can be used for amplifying various biomaterial samples, wherein different biomaterial samples comprise a male genome DNA extracted by a Chelex100 method, a magnetic bead extracting method or an organic extracting method and male blood or oral cells of human, which is/are collected by any one carrier of filterpaper, an FTA card, a cotton bud, gauze and the like. The kit has higher amplification specificity and higher thermal tolerance.

The invention discloses a method for simultaneously detecting twelve kinds of common respiratory viruses. According to the method, primers and probes are designed according to gene conservative areas of the twelve kinds of common respiratory viruses, namely influenza A virus, influenza B virus, influenza C virus, parainfluenza virus type 1, parainfluenza virus type 2, parainfluenza virus type 3, rhinovirus, Bocavirus, adenovirus, coronavirus, metapneumovirus and respiratory syncytial virus, nucleic acid fragments of samples to be measured are extracted for amplifying, and finally, the samples are separated by using a capillary electrophoresis method. The method disclosed by the invention has the advantages of low required sample size, high sensitivity and accuracy, good specificity and low cost; the defects that the conventional single tube multiplex fluorescence PCR (Polymerase Chain Reaction) detection primers are difficult to design, and multicolor fluorescence mutually intervenes and is not easy to part are overcome, the defects that a chip detection method is tedious in operation, high in detection cost and the like are also overcome, and a new method is provided for screening the respiratory viruses.