Abstract: Methods and kits for detection and quantification of EGFRvIII in the peripheral blood for monitoring the therapy of a GBM patients.

Abstract: Methods and kits for joining fragmented nucleic acid sequences together are provided, including performing an amplifying step including contacting a sample suspected of including a fragmented target nucleic acid with a pair of external primers and a pair of self-complementary internal primers, and generating a full length target nucleic acid. The methods can include performing an amplifying step, a hybridizing step, and a detecting step. Furthermore, kits are provided that are designed for the detection of a target nucleic acid sequence.

Abstract: Provided herein is technology relating to the amplification-based detection of nucleic acids and particularly, but not exclusively, to methods and compositions for minimizing variability in the activity between different samples or manufacturing lots of DNA polymerases, such as Taq DNA polymerase.

Abstract: Methods for detecting HAV in a biological sample are provided, comprising amplifying a target nucleic acid comprising the sequence of HAV in a reaction mixture. The reaction mixture comprises a biological sample which may contain the target nucleic acid and set of oligonucleotides. The invention also provides kits for the detection of HAV.

Abstract: A fluidic testing system is presented that includes a plurality of test chambers, a plurality of inlet channels, and a fluidic network that connects the inlet channels to one or more other chambers. The plurality of test chambers are each characterized by a length and a hydraulic diameter. The length of each test chamber is aligned substantially parallel to a gravity vector. Each of the test chambers has only one opening disposed along the length of the corresponding test chamber. Additionally, each of the test chambers is coupled via its respective opening to only one of the plurality of inlet channels.

Abstract: A fluidic testing method is presented that includes flowing an initial amount of liquid down a first inlet channel of a single-port fluidic testing system, and splitting the initial amount of liquid from the first inlet channel into a plurality of second inlet channels. Each second inlet channel is coupled to a given test chamber of a plurality of test chambers, wherein each of the plurality of test chambers has a wall that defines a longest side of a given test chamber, and wherein each of the plurality of test chambers has only one opening through the wall of a corresponding test chamber. The method also includes filling each of the plurality of test chambers with a final amount of liquid, the final amount being substantially equal in each of the test chambers and summing from each test chamber to equal the initial amount of liquid.

Abstract: The present invention provides methods of amplifying a target nucleic acid utilizing a circularized template. Circularization may be achieved utilizing a bridging oligonucleotide or an inverter primer. The bridging oligonucleotide or inverted primer is extended forming a concatemeric amplicon that can then be used as a template to provide exponential amplification of the target nucleic acid.

Abstract: Methods, compositions, and systems are provided that allow for reliable sequencing of the initial sequence region of a sequence of interest. The methods of the invention allow for more reliable barcoding of subpopulations of nucleic acids to be sequenced.

Abstract: A method of quantifying nucleic acids in a sample includes generating a plurality of droplets in oil within a microfluidic device, wherein at least some of the droplets comprise a nucleic acid, amplification reagents, and a fluorescent probe or dyes contained therein. The droplets are delivered to a collection chamber to form an array of droplets. The droplets are subject to thermal cycling within the collection chamber a plurality of times to perform nucleic acid amplification within the droplets. The array of droplets is imaged during the plurality of thermal cycles as well as at a thermal cycle endpoint. An initial concentration of nucleic acid in the sample is calculated based on at least one of: a ratio of aqueous phase droplets exhibiting fluorescence within the array at the thermal cycle endpoint or a cycle threshold (Ct) of one or more aqueous phase droplets within the array.

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: A method of amplifying a telomere of genomic DNA using an adaptor sequence, and a composition and a kit for amplifying the telomere of genomic DNA.

Abstract: The present invention is directed to methods and kits for gene analysis. The methods of the invention comprise the steps of providing nucleic acid; synthesis of a single-stranded DNA that is complementary to said nucleic acid molecule by contacting the nucleic acid with a DNA polymerase, a primer and a mixture of dNTPs under conditions that allow the generation of the DNA, wherein the primer comprises a target-complementary region and wherein the dNTP mixture comprises dATP, dGTP, dCTP, dTTP and dUTP; cleaving the DNA 5? to dU sites by (i) contacting the DNA with an uracil deglycosylase to generate a basic sites at positions of dUTP incorporation in the DNA; and (ii) contacting the DNA with an apurinic/apyrimidinic (AP) endonuclease; contacting the DNA comprising at its 5?-end the nucleotide sequence of the primer with a ssDNA ligase to circularize the DNA; and sequencing the circularized cDNA. The kits comprise the components necessary to perform the methods of the invention.

Abstract: Provided herein are systems, devices, and methods for generating thermal gradients in channels and uses thereof. In particular, provided herein are system, methods, and devices employing first and second thermal layers positioned around a channel in order to create a thermal gradient across a cross-section of the channel having, for example, a nucleic acid denaturation zone, a nucleic acid annealing zone, and a nucleic acid polymerization zone. Such devices find use in, for example, nucleic acid amplification procedures, including digital polymerase chain reaction (dPCR) to temperature cycle droplets for amplification of nucleic acid templates within the droplets.

Abstract: A system and method for eluting a sample from a swab are presented. The system includes a chamber dimensioned to receive at least a portion of the length of at least one swab, a fluidic channel connected to the chamber, and an actuator. The chamber has at least one wall being a flexible film. The fluidic channel is configured to at least one of introduce and expel liquids from the chamber. The actuator is configured to contact an outer surface of the flexible film such that movement of the actuator against the outer surface of the flexible film causes a respective movement of the at least one swab when the at least one swab is disposed next to an inner surface of the flexible film. The respective movement of the at least one swab elutes the sample from the at least one swab into the chamber.

Abstract: The present system provides novel methods and compositions for selecting a particular strand of RNA and/or producing a cDNA library that results in an unbiased representation of RNA in a sample.

Abstract: The present disclosure relates to a sample assessment device. By way of example, the sample assessment device may include a substrate including a sample application region; an amplification region comprising a plurality of amplification reagents; a waste region comprising an entrance fluidically coupled to the amplification region and extending away from the amplification region; and a detection region spaced apart from the amplification region. The sample assessment device may also include a valve coupled to the substrate and configured to separate the amplification region from the detection region in a closed configuration, wherein the amplification region and the valve are positioned on the sample assessment device between the sample application region and the detection region and wherein the sample assessment device is configured to permit lateral flow from the amplification region to the detection region when the valve is in an open configuration.

Abstract: Disclosed are methods for detecting a target nucleic acid in sample, which include contacting the sample with an oligonucleotide having a hairpin structure, where the oligonucleotide includes, in the 5? to 3? direction, an arbitrary sequence, an endonuclease recognition site for a nicking reaction, a sequence complementary to the arbitrary sequence, and a sequence complementary to the target nucleic acid; a polymerase; and an endonuclease capable of nicking the endonuclease recognition site. The disclosure also provides compositions and kits comprising an oligonucleotide having a hairpin structure, where the oligonucleotide includes, in the 5? to 3? direction an arbitrary sequence, an endonuclease recognition site for a nicking reaction, a sequence complementary to the arbitrary sequence, and a sequence complementary to a target nucleic acid.

Abstract: The present invention provides methods for the amplification of nucleic acid molecules. Methods for amplifying target polynucleotides, including mRNA, using oligonucleotides, DNA and RNA polymerases are provided. The invention further provides compositions and kits for practicing the methods, as well as methods which use the amplification products.

Abstract: Two genes required for this mercury methylation have been identified in bacteria and archaea. These genes are the hgcA gene and hgcB, a corrinoid protein that facilitates methyl group transfer to Hg, and a corrinoid protein-associated ferredoxin with two [4Fe-4S] binding motifs involved in generating cob(I)almin, respectively. The invention provides nucleic acid probes and primers for detecting methylmercury and or for assessing mercury methylation potential in environmental, clinical and other samples. The invention also provides antibodies against these proteins, antibodies against these proteins, methods of using the antibodies and methods of biocatalysis.

Abstract: This disclosure concerns a system and method for detecting heterologous DNA in plant materials.