Abstract: Described herein are products and processes for nucleic acid quantification, which are in part useful for detecting and determining the nucleotide sequence of rare nucleic acids (i.e., low copy number nucleic acids) in a sample. Such products and processes are useful for reducing the dynamic range among different nucleic acid species.

Abstract: In some embodiments, the present disclosure pertains to new compositions of matter that comprise phosphorescent reporters. In some embodiments, the phosphorescent reporters of the present disclosure comprise strontium aluminate. In some embodiments, the strontium aluminate is doped with europium and dysprosium (SrAl2O4:Eu2+, Dy3+). Additional embodiments of the present disclosure pertain to methods of making the aforementioned phosphorescent reporters. In some embodiments, the method includes size reduction of inorganic phosphorescent powders through a combination of wet milling and settling. In additional embodiments, the present disclosure pertains to methods of detecting the phosphorescent reporters in various settings, such as diagnostic settings.

Abstract: In a method and arrangement for detecting binding events of molecules, at least one second molecule covalently bound to a ligand, and optionally bonded to a third molecule bound to a ligand, forms a specific non-covalent fond with a first molecule immobilised on a bioactive surface by a covalent bond, and binding events between an analyte molecule and the second-molecule bound ligand are detected by an analytical measuring method, wherein the specific non-covalent bond between the first and second molecules is breakable by supplying a buffer solution, and, hence, a specific ligand, or two, different specific ligands are made available on the bioactive surface in an easily reversible manner.

Abstract: Novel methods and compositions for identifying one or more factors associated with a nucleic acid sequence (e.g., DNA and/or RNA) of interest are provided.

Abstract: A method of sequencing nucleic acids is provided using sequencing by ligation and/or sequencing by hybridization.

Abstract: Methods of directly isolating nucleic acid from an embedded biological sample are provided. An emulsified digest is generated in the presence of a thermostable protease, and an additive selected from an alkylene glycol, a poly(alkylene glycol), or a block copolymer having an average Mn of 76 to 2900, or a salt or derivative or combination thereof. Nucleic acid is isolated directly from the emulsified digest. The methods eliminate the use of organic solvents such as xylene in a deparaffinization step prior to isolating nucleic acids from paraffin-embedded samples, for example.

Abstract: Methods of identifying DNase I Hyper-Resistant Sites (DHRS), or in board sense, highly compact chromatin and characterizing the DNA methylation status of DMRs such as CpG islands and CpG island shores are provided. The methods are particularly useful for analysis of genomic DNA from low quantities of cells, for example, less than 1,000 cells, less than 100 cells, less than 10 cells, or even one cell, and can be used to generate chromatin and methylation profiles. The downstream analyses include in parallel massive sequencing, microarray, PCR and Sanger sequencing, hybridization and other platforms. These methods can be used to generate chromatin and DNA methylation profiles in drug development, diagnostics, and therapeutic applications are also provided.

Abstract: A method is provided herein, the method includes: applying a sample comprising target nucleic acids to a sample application zone of a substrate; and flowing a nucleic acid amplification reaction mixture across a length of the substrate through the sample application zone to amplify the target nucleic acid forming a nucleic acid amplification product; wherein the target nucleic acid having a first molecular weight is substantially immobilized at the sample application zone and wherein the amplification product having a second molecular weight migrates away from the sample application zone. An associated device is also provided.

Abstract: This invention generally relates to particle-assisted nucleic acid sequencing. In some embodiments, sequencing may be performed in a microfluidic device, which can offer desirable properties, for example, minimal use of reagents, facile scale-up, and/or high throughput. In one embodiment, a target nucleic acid may be exposed to particles having nucleic acid probes. By determining the binding of the particles to the target nucleic acid, the sequence of the target nucleic acid (or at least a portion of the target nucleic acid) can be determined. The target nucleic acid may be encapsulated within a fluidic droplet with the particles having nucleic acid probes, in certain instances. In some cases, the sequence of the target nucleic acid may be determined, based on binding of the particles, using sequencing by hybridization (SBH) algorithms or other known techniques.

Abstract: Disclosed herein are hybridization buffer compositions and hybridization compositions, methods of making the compositions, and methods of using the compositions, such as the hybridization of DNA or RNA sequences by fluorescence in situ hybridization (“FISH”).

Abstract: Methods for typing a strain of an organism are provided, the methods comprising the steps of amplifying, in a single reaction mixture containing nucleic acid from the organism, dividing the reaction mixture into a plurality of sets of second-stage reaction wells, each set of second-stage reaction wells containing a different pair of second-stage primers, subjecting each of the second-stage reaction wells to amplification conditions to generate a plurality of second-stage amplicons, melting the second-stage amplicons to generate a melting curve for each second-stage amplicon, and identifying the strain of the organism from the melting curves.

Abstract: The present invention relates to in situ hybridization methods for detecting a target nucleic acid in a biological sample comprising performing one or more method steps (e.g., pretreatment, denaturation, hybridization, washes) at room temperature. The invention further relates to kits for performing such methods.

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: The invention generally relates to compositions for maximizing capture of affinity-labeled molecules on solid supports. The disclosed methods and compositions were developed to maximize depletion of ribosomal RNA from total RNA samples, which is useful to improve the quality of RNA preparations used for applications such as massively parallel sequencing. The RNA depletion method is based on using long affinity-labeled RNA molecules that are complementary to all or part of the target ribosomal RNAs, as subtractive hybridization probes.

Abstract: Methods and kits for preparing nucleic acid fragments from a sample of purified nucleic acid are provided. Alternatively, chromatin or other long polymers can be sheared with similar methods and kits.

Abstract: Provided herein is a method for sequencing a polynucleotide molecules. The method includes the steps of providing a plurality of polynucleotide molecules attached to a surface, wherein a first portion of each polynucleotide molecule is attached to a first location of the surface and a second portion of each polynucleotide molecule is attached to a second location of the surface, the relative proximity of the first and second locations being correlated with the probability that the first and second portions are paired, separating the first and second portions of the polynucleotide molecules on the surface, determining the sequences of the first and second portions of the polynucleotide molecules and comparing the relative proximities and the sequences to determine which first and second portions are paired and to determine the sequence of the target polynucleotide molecules.

Abstract: Engineered nucleases (e.g., zinc finger nucleases (ZFNs), transcriptional activator-like effector nucleases (TALENs), and others) are promising tools for genome manipulation and determining off-target cleavage sites of these enzymes is of great interest. We developed an in vitro selection method that interrogates 1011 DNA sequences for their ability to be cleaved by active, dimeric nucleases, e.g., ZFNs and TALENs. The method revealed hundreds of thousands of DNA sequences, some present in the human genome, that can be cleaved in vitro by two ZFNs, CCR5-224 and VF2468, which target the endogenous human CCR5 and VEGF-A genes, respectively. Our findings establish an energy compensation model of ZFN specificity in which excess binding energy contributes to off-target ZFN cleavage and suggest strategies for the improvement of future nuclease design. It was also observed that TALENs can achieve cleavage specificity similar to or higher than that observed in ZFNs.

Abstract: This disclosure provides a method comprising: a) clamping the top and bottom strands of a double stranded DNA molecule to produce a duplex in which the top and bottom strands are linked; b) denaturing the duplex to produce a denatured product; and c) renaturing the denatured product in the presence of a labeled oligonucleotide that is complementary to a sequence of nucleotides in the double stranded DNA molecule, thereby producing a D-loop-containing product. Kits for performing the method and products made by the method are also provided.

Abstract: A method of identifying a first target nucleic acid comprising, providing a sample comprising the first target nucleic acid, providing a first set of paired oligonucleotides with complementarity to the first target nucleic acid, the first set of paired oligonucleotides comprising a first ratio of (a) first competitive oligonucleotides to (b) first signal oligonucleotides comprising a signal tag, wherein the competitive oligonucleotides compete with the signal oligonucleotides for binding to the first target nucleic acid, amplifying the first target nucleic acid with the polymerase chain reaction, thereby degrading the first signal oligonucleotide and permitting generation of a first signal, generating the first signal, measuring intensity of the first signal, and correlating the intensity of the first signal to the first ratio, thereby identifying the first target nucleic acid.

Abstract: The present invention relates to a method for analyzing biomolecules by using an oligonucleotide, and specifically, provides a method and an apparatus for determining the meaning of biomolecules in a biosample by analyzing through one examination of the biomolecules contained in the biosample by preparing, with respect to a nucleic acid including a target nucleic acid such as nucleic acid and receptor-analysis ligand conjugates, an oligonucleotide, which complementarily binds completely to a specific region of the target nucleic acid, and analyzing the target nucleic acid.