Abstract: Loop-shaped primer used in nucleic acid amplification is an oligonucleotide with 3-20 bases in both 3? and 5? ends which can be combined together to form a double-strand under appropriate conditions, resulting in the primer forming a stem-loop structure. The double-stranded structure is opened and the stem-loop structure dissolves when the primer recognizes and hybridizes with the target sequence. If the target sequence is not present the primer can form a stem-loop structure automatically by self-annealing. The primer can comprise a universal tag sequence or not. Together with universal tag sequence primer the primer comprising the universal tag sequence can be used for a second round of amplification. The primer has high specificity and does not form a primer dimmer. The primer is easy to design and is suitable for measuring gene expression and detecting features of nucleic acids such as SNPs and rare mutations.

Abstract: Provided herein are methods for detecting mutations in nucleic acid, and compositions and kits for performing such methods. In particular, nucleic acid amplification and fluorescence detection methods are provided to detect mutations and assess the mutational load. The methods are based on a set of adjacently binding probes wherein one probe is labelled with a quencher and the other is a self-indicating probe labelled with fluorophore and quencher. The binding of the probes is analyzed by melting curve analysis.

Abstract: Individual temperature control in multiple reactions performed simultaneously in a spatial array such as a multi-well plate is achieved by thermoelectric modules with individual control, with each module supplying heat to or drawing heat from a single region within the array, the region containing either a single reaction vessel or a group of reaction vessels.

Abstract: The present invention provides methods for analyzing large nucleic acids including chromosomes and chromosomal fragments. In one aspect, the present invention provides a method of nucleic acid analysis comprising the steps of (a) obtaining a sample of nucleic acid comprising at least one chromosome or fragment greater than about 1000 base pairs in length and containing a target region; (b) creating an emulsion in which each drop of the emulsion contains an average of between about 0-2, 0-1.75, 0-1.5, 0-1.0, 0-0.75, 0-0.5, or fewer chromosomes or fragments of step (a), (c) performing emulsion PCR, (d) quantifying the number of emulsion droplets containing amplified nucleic acid from the target region; (e) calculating the ratio of droplets containing amplified nucleic acid from the target region to total droplets; and (f) comparing the ratio of step (e) to a reference ratio representing a known genotype.

Abstract: The present invention provides a reaction vessel (20) for a PCR device. The reaction vessel (20) comprises a sample vial (32) defining a reaction chamber (33) for performing PCR and a storage vessel (62) defining a storage chamber (63) for optical detection. The reaction chamber (33) is in fluid communication with a liquid supply port (34) for supplying a liquid sample containing at least one target DNA to the reaction chamber (33). The reaction chamber (33) and the storage chamber (63) are in fluid communication via a spacer element (42) and a porous membrane (51) for hybridization of the at least one target DNA within the liquid sample onto specific immobilised hybridization probes. The lower end of the spacer element (42) extends into the reaction chamber (33), but does not reach the bottom thereof. The upper end of the spacer element (42) is located in proximity of the porous membrane (51), which is made from a material having different physical properties in a dry state and a wet state.

Abstract: In alternative embodiments, the invention provides computational algorithms, computer programs, software and other methods, systems and products of manufacture (e.g., computers, devices or apparatus) for identifying members of microbial communities, their abundance and distribution from amplicon sequence data, and comparing microbial communities and consortia. In alternative embodiments, the invention provides computer-implemented methods comprising a subset of, substantially all, or all of the steps as set forth in the flow chart of FIG. 1, FIG. 3 or FIG. 4. In alternative embodiments, the invention provides methods for identification of consortia, optionally followed by construction of artificial microbial consortia from pure strains or enrichment cultures. In alternative embodiments, the invention provides compositions, fluids, bioreactors, muds, reservoirs or products of manufacture comprising a synthetic microbial consortium made by the method of the invention.

Abstract: The present invention relates to systems and methods for monitoring the amplification of DNA molecules and the dissociation behavior of the DNA molecules. A method according to one embodiment of the invention may include the steps of: forcing a sample of a solution containing real-time PCR reagents to move though a channel; and while the sample is moving through an analysis region of the channel, performing the steps of: (a) cycling the temperature of the sample until the occurrence of a predetermined event; (b) after performing step (a), causing the sample's temperature to gradually increase from a first temperature to a second temperature; and (c) while the step of gradually increasing the sample's temperature is performed, using an image sensor to monitor emissions from the sample.

Abstract: Methods and kits for DNA repair are provided. The methods and kits described herein repair multiple types of DNA damage. The kit may include a plurality of enzymes to repair a greater variety of lesions than any single enzyme is capable of repairing. Repair of damaged DNA may include releasing damaged bases from the DNA strand, nicking the DNA at the damaged sites, translating the nicks via 5?-3? exonuclease activity, and sealing the nicks. The enzymes employed in the repair process may then be heat-inactivated, thereby obviating a purification process. The repaired DNA may then be analyzed using a variety of DNA analysis methods.

Abstract: A multiplex polymerase chain reaction assay that targets nine tetranucleotide short tandem repeat (STR) markers in the mouse genome. Unique profiles were obtained from seventy-two mouse samples that were used to determine the allele distribution for each STR marker. Correlations between allele fragment length and repeat number were determined with DNA Sanger sequencing. Genotypes for L929 and NIH3T3 cell lines were shown to be stable with increasing passage numbers as there were no significant differences in fragment length with samples of low passage when compared to high passage samples. In order to detect cell line contaminants, primers for two human STR markers were incorporated into the multiplex assay to facilitate detection of human and African green monkey DNA. This multiplex assay is the first of its kind to provide a unique STR profile for each individual mouse sample and can be used to authenticate mouse cell lines.

Abstract: An apparatus for performing a Polymerase Chain Reaction (PCR) is disclosed. The apparatus comprises a PCR chamber for performing a Polymerase Chain Reaction and a printed circuit board (PCB) fluidic device. The PCR chamber is a fluidic chamber and is located in, or is part of, the printed circuit board (PCB) fluidic device. A method for manufacturing an apparatus for performing the Polymerase Chain Reaction and a method for performing the Polymerase Chain Reaction are further disclosed.

Abstract: The method of the invention comprises the stratification of a cancer patient population into various cancer therapy groups based on analysis by genomic DNA microarray of multiple gene amplifications or deletions present or absent in the diseased tissue of each patient. In particular, the invention involves patient stratification into one of at least four cancer therapy groups based on the microarray analysis of gene amplification or gene deletion at multiple chromosome locations.

Abstract: Oligonucleotide primer useful for synthesizing a cDNA copy of HIV-1 nucleic acids from a broad range of HIV-1 subtypes, including M group and O group variants.

Abstract: A method and apparatus are provided for identifying a biological sample obtained during either paternity screening, genetic screening, prenatal diagnosis, presymptomatic diagnosis, diagnosis to detect the presence of a target microorganism carrier detection analysis, forensic chemical analysis, or diagnosis of a subject to determine whether a subject is afflicted with a particular disease or disorder, or is at risk of developing a particular disorder, wherein the result obtained from the analysis is associated with the unique DNA fingerprint biological barcode of the genotype of the subject being analyzed. The methods and apparatus of the invention have application in the fields of diagnostic medicine, disease diagnosis in animals and plants, identification of genetically inherited diseases in humans, family relationship analysis, forensic analysis, and microbial typing.

Abstract: Universal reference dye mixtures for quantitative amplification, and uses thereof, are provided.

Abstract: The present invention relates to methods for identifying compounds that modulate untranslated region-dependent expression of a target gene. The invention particularly relates to using untranslated regions of a target gene or fragments thereof linked to a reporter gene to identify compounds that modulate untranslated region-dependent expression of a target gene. The methods of the present invention provide a simple, sensitive assay for high-throughput screening of libraries of compounds to identify pharmaceutical leads.

Abstract: The present invention provides methods, kits, and compositions for producing single-stranded circular DNA by PCR. In particular, hairpin primers are provided, and methods of use thereof to produce single-stranded circular DNA molecules.

Abstract: A reactor for the quantitative analysis of target nucleic acids using an evanescent wave detection technique and a method of use thereof is provided. The reactor includes a substrate with a cavity, a buffer layer arranged over the substrate; a cover plate arranged over the buffer layer, and inlet and outlet ports. The reactor is thermally and chemically stable for PCR processing and suitable for an evanescent wave detection technique.

Abstract: The present invention provides methods and compositions for the detection of novel BRAF splice variants that mediate resistance to BRAF and/or pan-RAF inhibitors. In particular, the invention provides PCR primer(s) to be used in the disclosed methods of detection. In some embodiments, the compositions and methods of the present invention are used to predict resistance to BRAF and/or pan-RAF inhibitors in a subject suffering from or suspected of having cancer and further provides alternative treatment strategy(ies) for a subject predicted to be resistant to BRAF and/or pan-RAF inhibitors. In a further embodiment, methods and composition for the identification of novel agents useful to overcome resistance to BRAF and/or pan-RAF inhibitors are disclosed. The present invention also provides isolated polynucleotide sequences of novel 5? BRAF splice variant(s) and proteins produced from such polynucleotide sequences as well as cell line(s) that endogenously or exogenously express the splice variant(s).

Abstract: A non-symmetric polymerize chain reaction (PCR) amplification method employing a limiting primer in low concentration whose concentration-adjusted melting point at least equals, and preferably exceeds, that of the excess primer, the latter in turn not being more than 25° C. below the melting temperature of the amplicon. Assays employing such amplification and labeled hybridization probes, including assays that include a detection step following primer extension or a low-temperature probe, or both. Kits for performing such assays and primer or primer-and-probe sets for performing the foregoing amplifications and assays.

Abstract: Methods for sequencing single large DNA molecules by clonal multiple displacement amplification using barcoded primers. Sequences are binned based on barcode sequences and sequenced using a microdroplet-based method for sequencing large polynucleotide templates to enable assembly of haplotype-resolved complex genomes and metagenomes.