Abstract: The present invention relates generally to methods for generating single stranded nucleic acid molecules following enhanced solid phase polynucleotide amplification. The present invention employs an amplification reaction using primers with differential priming properties at particular annealing conditions or an immobilized primer nested between two aqueous phase primers. Thus, by primer design, solid support primer participation is enhanced relative to aqueous phase primers. The subject invention further provides methods for labeling solid matrices with single and double stranded nucleic acid molecules. Kits for generating single stranded nucleic acid molecules and for conducting amplification reactions also form part of the present invention. The present invention further provides amplification systems for the generation of single stranded nucleic acid molecules optionally labelled with a reporter molecule and their use inter alia as labels, primers and probes.

Abstract: Efficient methods are disclosed for the high throughput identification of mutations in genes in members of mutagenized populations. The methods comprise DNA isolation, pooling, amplification, creation of libraries, high throughput sequencing of libraries, preferably by sequencing-by-synthesis technologies, identification of mutations and identification of the member of the population carrying the mutation and identification of the mutation.

Abstract: The invention provides a method and apparatus for detecting a signal of a specific spectrum emitted in the course of a chemical or biochemical reaction. The method comprises conducting the reaction in a reaction vessel, which is arranged so that light emanating from the reaction vessel is received by a detector comprising a plurality of photosensors in an array, wherein each photosensor is activated by light falling within a particular waveband range only, and where photosensors activated by light in different waveband ranges are distributed throughout the array. Output from one or more subsets of those photosensors which receive wavebands which contribute to the said specific spectrum is monitored and the output from a subset, or the relationship between the outputs of each subset are used to determine the signal in the specific spectrum.

Abstract: The invention generally relates to droplet based digital PCR and methods for analyzing a target nucleic acid using the same. In certain embodiments, methods of the invention involve forming sample droplets containing, on average, a single target nucleic acid, amplifying the target in the droplets, excluding droplets containing amplicon from the target and amplicon from a variant of the target, and analyzing target amplicons.

Abstract: According to one embodiment, a multi-nucleic-acid amplification reaction tool includes a support and a plurality of types of primer sets. The support is configured to be able to support a reaction field of a liquid phase. A plurality of types of primer sets are fixed in a releasable state, for each type, on mutually independent fixing regions of at least a surface of the support, which is in contact with the reaction field, when the liquid phase forms the reaction field. A plurality of types of primer sets are configured to amplify the respectively corresponding target sequences.

Abstract: Provided herein are fluorescence detection methods for nucleic acid sequences and to kits for performing such methods.

Abstract: The present invention generally relates to methods and apparatuses for amplifying nucleic acid sequences using immobilized DNA polymerase. More particularly, it relates to methods and apparatuses useful for amplifying target nucleic acid sequences by forming a plurality of reaction regions in which polymerase chain reaction (PCR) can occur, positioning immobilized DNA polymerase in a specific reaction region, and circulating DNA through the reaction regions. The present invention provides those methods and apparatuses that allow simple separation and recovery of the DNA polymerase after the amplification, that can be operated not only with thermostable DNA polymerases but also with non-thermostable DNA polymerases, and that are simpler in their designs and processes so that they can be readily integrated into complex devices such as Lab-on-a-chip.

Abstract: Provided herein are methods, compositions, and kits for detecting alleles using a single probe or a single primer set. Also, provided herein are methods, compositions, and kits for detecting allelic variants using a single probe or a single primer set. Also, provided herein are methods, compositions, and kits for determining a polymerization error rate.

Abstract: The present invention provides systems, apparatuses, and methods to detect the presence of fetal cells when mixed with a population of maternal cells in a sample and to test fetal abnormalities, e.g. aneuploidy. The present invention involves labeling regions of genomic DNA in each cell in said mixed sample with different labels wherein each label is specific to each cell and quantifying the labeled regions of genomic DNA from each cell in the mixed sample. More particularly the invention involves quantifying labeled DNA polymorphisms from each cell in the mixed sample.

Abstract: Disclosed is a method for adjusting the amplification efficiency of a target polynucleotide in the amplification of the target polynucleotide by PCR using primers (i) to (iii) below, the method comprising adjusting the amplification efficiency of the target polynucleotide by changing the quantity ratio of the primers (i) to (iii) below: (i) a first primer which is able to be base-paired with the target polynucleotide; (ii) a second primer which is able to be base-paired with the target polynucleotide in competition with the first primer and from which extension reaction by PCR less occurs as compared to the first primer; and (iii) a third primer designed to allow for the amplification of the target polynucleotide in pairs with the first primer.

Abstract: The present disclosure provides an improved field effect transistor and device that can be used to sense and characterize a variety of materials. The field effect transistor and/or device including the transistor may be used for a variety of applications, including genome sequencing, protein sequencing, biomolecular sequencing, and detection of ions, molecules, chemicals, biomolecules, metal atoms, polymers, nanoparticles and the like.

Abstract: The present invention relates to a method for quantifying and/or detecting one or more nucleic acids of a genome in a sample, wherein in an amplification reaction, (i) a first nucleic acid is amplified, the locus that is amplified is a multicopy locus (MCL) within the genome, wherein the locus shares at least 80% sequence identity to a sequence according to SEQ ID NO. 1 over a stretch of 80 base pairs, and wherein the multicopy locus has copies on at least two different chromosomes, (ii) a second nucleic acid that has been added as an internal control (IC) is also amplified, and (iii) the amount of amplification product from the amplification of the first nucleic acid is determined.

Abstract: A method for replicating and amplifying a target nucleic acid sequence is described. A method of the invention involves the formation of a recombination intermediate without the prior denaturing of a nucleic acid duplex through the use of a recombination factor. The recombination intermediate is treated with a high fidelity polymerase to permit the replication and amplification of the target nucleic acid sequence. In preferred embodiments, the polymerase comprises a polymerase holoenzyme. In further preferred embodiments, the recombination factor is bacteriophage T4 UvsX protein or homologs from other species, and the polymerase holoenzyme comprises a polymerase enzyme, a clamp protein and a clamp loader protein, derived from viral, bacteriophage, prokaryotic, archaebacterial, or eukaryotic systems.

Abstract: A method for replicating and amplifying a target nucleic acid sequence is described. A method of the invention involves the formation of a recombination intermediate without the prior denaturing of a nucleic acid duplex through the use of a recombination factor. The recombination intermediate is treated with a high fidelity polymerase to permit the replication and amplification of the target nucleic acid sequence. In preferred embodiments, the polymerase comprises a polymerase holoenzyme. In further preferred embodiments, the recombination factor is bacteriophage T4 UvsX protein or homologs from other species, and the polymerase holoenzyme comprises a polymerase enzyme, a clamp protein and a clamp loader protein, derived from viral, bacteriophage, prokaryotic, archaebacterial, or eukaryotic systems.

Abstract: A method for replicating and amplifying a target nucleic acid sequence is described. A method of the invention involves the formation of a recombination intermediate without the prior denaturing of a nucleic acid duplex through the use of a recombination factor. The recombination intermediate is treated with a high fidelity polymerase to permit the replication and amplification of the target nucleic acid sequence. In preferred embodiments, the polymerase comprises a polymerase holoenzyme. In further preferred embodiments, the recombination factor is bacteriophage T4 UvsX protein or homologs from other species, and the polymerase holoenzyme comprises a polymerase enzyme, a clamp protein and a clamp loader protein, derived from viral, bacteriophage, prokaryotic, archaebacterial, or eukaryotic systems.

Abstract: Presented herein are methods and compositions for generating sequence-specific, secondary amplification products during Loop-mediated Isothermal Amplification (LAMP). Conventional LAMP produces a preponderance of high molecular weight DNA structures concatenated into self-complementary hairpins, which are not amenable to detection by routine probe-based hybridization methods, making multiplex detection of two or more targets or sequence variants in closed-tube formats extremely difficult. Provided herein, for example, are methods for generating secondary LAMP products bearing a fragment of the original target sequence embedded within low-molecular weight products that are devoid of competitive hairpin structures, the lack of which enhances probe-based detection of target sequences. These secondary products can, for example, be produced in real-time, during the LAMP process, and can provide the option of detecting multiple target sequences within a single tube using, e.g.

Abstract: The disclosure provides a composition and method for isolating nucleic acids, in which the composition includes at least one halocarbon, at least one salt and at least one surfactant. Mixing the composition and a biosample to form a homogenized solution, nucleic acids in the solution can be easily isolated with a simple treatment and good yield.

Abstract: The present invention concerns a method for a polymerase chain reaction, in which a template nucleic acid, at least one primer, deoxyribonucleoside triphosphates as well as a DNA polymerase with proofreading activity are used. In addition, according to this invention, at least one target substrate is added to the polymerase chain reaction, whereby the efficiency of the DNA polymerase with proofreading activity is significantly increased. Any molecule that reduces or, in the optimal case, blocks the 3?,5?-exonuclease activity of the DNA polymerase used is suitable as target substrate. Technical solutions for the added substrate (target substrate) are in particular single stranded, linear oligonucleotides, hairpin oligonucleotides and RNA and DNA molecules. Furthermore, a kit is disclosed which comprise the required reagents for the implementation of the method according to the invention.

Abstract: The invention relates to an optical detection system for a thermal cycling device including at least one light source, a light detection device for detecting light received from a plurality of biological samples, and a lens having first and second surfaces formed on the lens, the second surface substantially opposed to the first surface. The first surface may be configured to collimate light and the second surface may be configured to direct light into each of the plurality of biological samples.

Abstract: In some embodiments, the present teachings provide methods for nucleic acid amplification, comprising forming a reaction mixture, and subjecting the reaction mixture to conditions suitable for nucleic acid amplification. In some embodiments, methods for nucleic acid amplification include subjecting the nucleic acid to be amplified to partially denaturing conditions. In some embodiments, methods for nucleic acid amplification include amplifying without fully denaturing the nucleic acid that is amplified. In some embodiments, the methods for nucleic acid amplification employ an enzyme that catalyzes homologous recombination and a polymerase. In some embodiments, methods for nucleic acid amplification can be conducted in a single reaction vessel. In some embodiments, methods for nucleic acid amplification can be conducted in a single continuous liquid phase of a reaction mixture, without need for compartmentalization of the reaction mixture or immobilization of reaction components.

Abstract: The present disclosure provides methods for determining phased nucleic acid sequence for a single chromosome of interest and/or a single chromosomal fragment of interest. The present disclosure also provides methods for determining phased nucleic acid sequence for a plurality of single chromosomes of interest and/or a plurality of single chromosomal fragments of interest. The plurality of single chromosomes of interest may be of one chromosome type or of two or more chromosome types. The present disclosure also provides a method for isolating a plurality of chromosomal fragments of a specified size range, where the chromosomal fragments are from one or more specified regions of the genome. The plurality of chromosomal fragments may be separated into single chromosomal fragments and sequenced to provide phased nucleic acid sequence for the single chromosomal fragments.

Abstract: Methods for amplification and sequencing of at least one nucleic acid comprising the following steps: (1) forming at least one nucleic acid template comprising the nucleic acid(s) to be amplified or sequenced, wherein said nucleic acid(s) contains at the 5? end an oligonucleotide sequence Y and at the 3? end an oligonucleotide sequence Z and, in addition, the nucleic acid(s) carry at the 5? end a means for attaching the nucleic acid(s) to a solid support; (2) mixing said nucleic acid template(s) with one or more colony primers X, which can hybridize to the oligonucleotide sequence Z and carries at the 5? end a means for attaching the colony primers to a solid support, in the presence of a solid support so that the 5? ends of both the nucleic acid template and the colony primers bind to the solid support; (3) performing one or more nucleic acid amplification reactions on the bound template(s), so that nucleic acid colonies are generated and optionally, performing at least one step of sequence determination of

Abstract: Disclosed is a method for obtaining a bifunctional complex comprising a display molecule part and a coding part, wherein a nascent bifunctional complex comprising a chemical reaction site and a priming site for enzymatic addition of a tag is reacted at the chemical reaction site with one or more reactants, and provided with respective tag(s) identifying the reactant(s) at the priming site is using one or more enzymes.

Abstract: A method is disclosed for rapid molecular profiling of tissue or other cellular specimens by placing a donor specimen in an assigned location in a recipient array, providing copies of the array, and performing a different biological analysis of each copy. The results of the different biological analyses are compared to determine if there are correlations between the results of the different biological analyses at each assigned location. In some embodiments, the specimens may be tissue specimens from different tumors, which are subjected to multiple parallel molecular (including genetic and immunological) analyses. The results of the parallel analyses are then used to detect common molecular characteristic of the genetic disorder type, which can subsequently be used in the diagnosis or treatment of the disease. The biological characteristics of the tissue can be correlated with clinical or other information, to detect characteristics associated with the tissue.

Abstract: An attachment unit for attachment of a reaction container including a channel filled with a reaction solution containing a fluorescent probe and a liquid having a specific gravity different from that of the reaction solution and being immiscible with the reaction solution, the reaction solution moving close to opposed inner walls, a first heating unit heating a first region of the channel and a second heating unit heating a second region of the channel when the reaction container is attached to the attachment unit, a drive mechanism switching arrangement of the attachment unit, the first heating unit, and the second heating unit between a first arrangement and a second arrangement in which a lowermost position of the channel is located within a first region and a second region, respectively, a measurement unit measuring light intensity, and a control unit controlling the portions described above.

Abstract: Compositions, methods and kits for detecting Chikungunya viral nucleic acids. Particularly described are methods for detecting very low levels of the viral nucleic acids using nucleic acid amplification.

Abstract: A method for the detection of a target nucleic acid, which method comprises contacting template nucleic acid from a sample with (i) a signalling system and (ii) a tailed nucleic acid primer having a template binding region and the tail comprising a linker and a target binding region, in the presence of appropriate nucleoside triphosphates and an agent for polymerisation thereof, under conditions such that the template binding region of the primer will hybridise to a complementary sequence in the template nucleic acid and be extended to form a primer extension product, separating any such product from the template whereupon the target binding region in the tail of the primer will hybridise to a sequence in the primer extension product corresponding to the target nucleic acid, and wherein any such target specific hybridisation causes a detectable change in the signalling system, such that the presence or absence of the target nucleic acid in the sample is detected by reference to the presence or absence of a de

Abstract: Provided is a method of detecting or quantifying a wheat species-specific DNA in a test sample by polymerase chain reaction. The method comprises a step of amplifying a nucleic acid molecule having a partial sequence of a nucleotide sequence identified as SEQ ID NO: 1 using a nucleic acid molecule in the test sample or a nucleic acid molecule extracted from the test sample as the template and using a primer pair capable of amplifying the partial sequence and a step of detecting or quantifying the amplified nucleic acid molecule.

Abstract: A system (20) for a PCR reaction includes an array of reaction vessels mounted on a thermal mount (21). The thermal mount (21) is provided with a liquid path therein coupled to a cooling liquid input port (22), a heating liquid input port (23) and a liquid output port (24). A pump (38) is used to pump liquid from cooling liquid source (29) either along a cooling liquid path (28) to the cooling liquid input port (22), or via a heating liquid source (31), where the liquid is heated, and along a heating liquid path (30) to the heating liquid input port (23). A temperature sensor (34) measures the temperature of the thermal mount (21) and a processor (27) controls the pump, valves (26) at the input and output ports and valves (41-44) at either side of the pump (38), to control whether heating or cooling liquid is input to the thermal mount, and at what flow rate, in order to obtain the correct temperature of the thermal block (21).

Abstract: Polypeptides having nucleic acid binding activity are provided. Methods of using polypeptides having nucleic acid binding activity are provided. Fusion proteins and methods of using fusion proteins are provided. Fusion proteins comprising a polymerase and a nucleic acid binding polypeptide are provided. Fusion proteins comprising a reverse transcriptase and a nucleic acid binding polypeptide are provided. Methods are provided for amplifying a nucleic acid sequence using a fusion protein comprising a nucleic acid binding polypeptide and a polymerase. Methods are provided for amplifying a nucleic acid sequence using a fusion protein comprising a nucleic acid binding polypeptide and a reverse transcriptase.

Abstract: The purpose of the subject invention is to provide a method of manufacturing a mixture of amplified double-stranded nucleic acids comprising unknown sequence including the complete 5? end sequence. A method of manufacturing a mixture of amplified double-stranded nucleic acids comprising: (a) preparing a single-stranded nucleic acid comprising a single-stranded adapter 1, a single-stranded nucleic acid fragment and a single-stranded adapter 2, and (b) conducting PCR with said single-stranded nucleic acid prepared in step (a), a primer 1, and a primer 2 to amplify double-stranded nucleic acids.

Abstract: The present invention relates to methods for screening of reagents used in the performance of polymerase chain reaction (PCR) assays. The invention has applications for genotyping, pathogen detection and in vitro diagnostics.

Abstract: The technology described herein generally relates to microfluidic cartridges configured to amplify and detect polynucleotides extracted from multiple biological samples in parallel. The technology includes a microfluidic substrate, comprising: a plurality of sample lanes, wherein each of the plurality of sample lanes comprises a microfluidic network having, in fluid communication with one another: an inlet; a first valve and a second valve; a first channel leading from the inlet, via the first valve, to a reaction chamber; and a second channel leading from the reaction chamber, via the second valve, to a vent.

Abstract: Provided are disposable, moisture-activated, self-heating cartridges useful for, e.g., isothermal nucleic acid amplification, incubation, and thermal actuation, and visual fluorescent detection of the amplification products. These devices may be self-contained and do not require any special instruments to operate.

Abstract: The invention provides a method for dispensing liquid, comprising the steps of: (a) positioning a droplet to be dispensed in a gap of an electrowetting device using an electrowetting array; and (b) dispensing the droplet through a hole in a housing or substrate of the electrowetting device. The invention further provides a method for withdrawing liquid comprising the steps of: (a) positioning a droplet to be withdrawn from a gap of an electrowetting device using an electrowetting array; and (b) withdrawing the droplet through a hole in a housing or substrate of the electrowetting device.

Abstract: The invention provides a method of determining the nucleotide sequence of a target nucleic acid using a reversibly terminating nucleotide that is modified at the 2? position.

Abstract: A thermal cycler includes: a first mounting unit and a second mounting unit which are cylindrical; a temperature gradient forming unit which forms a temperature gradient along mounting directions of the first mounting unit and the second mounting unit; and a driving mechanism which rotates the first mounting unit, the second mounting unit, and the temperature gradient forming unit around a rotating shaft having a component perpendicular to a direction in which gravity is applied and a component intersecting the mounting directions of the first mounting unit and the second mounting unit, wherein the first mounting unit and the second mounting unit are disposed on opposite sides to each other with the rotating shaft interposed therebetween, the mounting direction of the first mounting unit and the mounting direction of the second mounting unit are in the same direction.

Abstract: Described are reagents, methods, and kits for eluting, and amplifying and/or characterizing DNA from liquid and dried blood samples. A one-step DNA elution buffer has been developed that simplifies purification of DNA from blood samples. The purified DNA is suitable for use in subsequent widely used techniques such as enzymatic DNA amplification and quantitative analysis such as real-time PCR.

Abstract: A two-step multiplex amplification reaction includes a first step which truncates the standard initial multiplex amplification round to “boost” the sample copy number by only a 100-1000 fold increase in the target. Following the first step the product is divided into optimized secondary single amplification reactions, each containing one of the primer sets that were used previously in the first or multiplexed booster step. The booster step can occur using an aqueous target nucleic acid or using a solid phase archived nucleic acid. In particular, nucleic acid sequences that uniquely identify E. Coli were identified using the multiplex amplification method.

Abstract: The invention is directed to a method for isothermally amplifying a DNA sequence involving hybridizing a destabilizing DNA template to complementary nucleotide fragments to form a first nicked duplex; ligating the first nicked duplex to form a product duplex comprising the DNA sequence and the template, wherein the product duplex is capable of dissociating to release the DNA sequence and the template; and repeating these steps to generate multiple copies of the template and the DNA sequence. Further, the method may also involve hybridizing the DNA sequence to complementary destabilizing fragments or probes to form a second nicked duplex; ligating the second nicked duplex to form the product duplex comprising the DNA sequence and the template, wherein the product duplex dissociates to release the DNA sequence and the template; and repeating these steps to generate multiple copies of the template and the DNA sequence.

Abstract: Provided herein are methods and compositions for the prognostic evaluation of a patient suspected of having, or having, cancer by assessing the expression of IMP3 in a biological sample of a patient. Methods can be used at the time of initial diagnosis of malignant tumors to identify a group of patients with a high potential to develop progression or metastasis later. Therefore, methods not only are able to provide very useful prognostic information for patients but also can help clinicians to select a candidate patient likely to benefit from early and aggressive cancer therapy. Methods and compositions for the treatment of cancer associated with expression of IMP3 are also provided.

Abstract: Primer sets for amplifying target regions containing sites to be detected in the UGT1A1 gene by a gene amplification method are provided, wherein the primer sets can amplify the regions specifically Three pairs of primer sets are used including forward primers consisting of the base sequences of SEQ ID NOs: 4 or 81, 21, and 42 as well as reverse primers consisting of the base sequences of SEQ ID NOs: 13 or 91, 29 and 48, respectively. The use of these primer sets makes it possible to amplify three target regions including parts where three types of polymorphisms (UGT1A1*6, UGT1A1*27, and UGT1A1*28) of the UGT1A1 gene are generated, respectively, in the same reaction solution at the same time.

Abstract: Compositions, reaction mixtures, and methods for performing an amplification reaction, including multiplex amplification reaction, wherein the method comprises using one or more amplification oligomer complexes comprising linked first and second amplification oligomer members. In one aspect, the amplification oligomer complex is hybridized to a target nucleic acid, the target nucleic acid with hybridized amplification oligomer complex is then captured, and other components are washed away. Target sequences of the target nucleic acids are pre-amplified to generate a first amplification product. The first amplification product is amplified in one or more secondary amplification reactions to generate second amplification products.

Abstract: The present invention provides methods to obtain dry compositions of reaction compounds that maintain the biological activity of the compounds upon re-solubilization after a certain storage time. Preferably, the dry composition comprises a polymerase, and the dry composition is usable for polymerase chain reaction (PCR) amplification after re-solubilization.

Abstract: Provided is a polynucleotide including, from the 3? terminus of the polynucleotide to the 5? terminus of the polynucleotide, a first region including a nucleotide sequence complementary to a nucleotide sequence of a portion of a target nucleic acid; a second region including a nucleotide sequence identical to a nucleotide sequence of a portion of the target nucleic acid; and a third region including a nucleotide sequence that self-hybridizes to form a stem-loop structure, and compositions, kits, and methods related thereto.

Abstract: An M×N matrix microfluidic device for performing a matrix of reactions, the device having a plurality of reaction cells in communication with one of either a sample inlet or a reagent inlet through a via formed within an elastomeric block of the device. Methods provided include a method for forming vias in parallel in an elastomeric layer of an elastomeric block of a microfluidic device, the method comprising using patterned photoresist masks and etching reagents to etch away regions or portions of an elastomeric layer of the elastomeric block.

Abstract: This document provides methods and materials for detecting contaminated food products. For example, methods and materials for using an enzymatic amplification cascade of restriction endonucleases to detect nucleic acid of a microorganism or virus (e.g., a pathogen) within a sample (e.g., food product sample) being tested, thereby assessing a food product for possible contamination are provided.

Abstract: Provided herein are methods and materials for diagnosing a subject's predisposition for pulmonary infection in a CF subject by detecting a pulmonary infection genetic marker. Pulmonary infection markers have been identified in the IL-1 gene cluster and may be useful in predicting CF disease progression and assessing a CF subject's response to therapy.

Abstract: Methods for performing a direct enzymatic reaction involving a nucleic acid molecule include performing the enzymatic reaction directly using a biological specimen in a reaction mixture containing a zwitterionic buffer and/or a non-reducing carbohydrate to prevent the biological specimen from inhibiting the enzymatic reaction, in which a nucleic acid molecule present in the biological specimen is not purified before the enzymatic reaction, and obtaining a product from the enzymatic reaction.

Abstract: Compositions, methods and kits for detecting viral nucleic acids. Targets that can be detected in accordance with the invention include HBV and/or HIV-1 and/or HCV nucleic acids. Particularly described are oligonucleotides that are useful as hybridization probes and amplification primers that facilitate detection of very low levels of HBV nucleic acids.