Abstract: The invention provides methods of purifying one or more biological analytes from a sample. The method comprises incubating the sample with nanoparticles and centrifuging a biphasic system. The biological analytes can be one or more of an enzyme, a protein, a nucleic acid, an organelle, a cell, a bacteria, a virus, or any other biological material which requires purification.

Abstract: A simple, rapid, inexpensive, and promising commercial biomarker assay method for multiple diseases is described herein. The present invention detects miRNA-based biomarkers in human stool specimens. The method of the present invention amplifies miRNA directly from stool specimens without any prior miRNA extraction. Differential expression of specific microRNAs in stool of colorectal cancer CRC and adenoma patients suggest fecal microRNAs as a novel potential biomarker for colorectal neoplasia detection. The method of the present invention has diagnostic, prognostic, and therapeutic relevance for gastroenterological cancers/colorectal cancer and as well as further acquired or hereditary GI diseases.

Abstract: An object of the present invention is to stimulate a T cell without using a peptide/MHC tetramer. In the present invention, the step of supplying an antigen peptide to a T cell having a T cell receptor (TCR) that can recognize the antigen peptide on cell surface to form a complex of a major histocompatibility complex (MHC) molecule on the cell surface of the T cell and the antigen peptide is used, and the T cell is stimulated through recognition by TCR of the antigen peptide as the MHC molecule-antigen peptide complex on the cell surface of the same T cell. Such a stimulating and activating method would be applicable to not only T cells, but also various cells. According to the present invention, an antigen-specific T cell can be identified without establishing any antigen-specific T cell strain, and without using such a reagent as MHC/peptide tetramer. That is, a cancer-specific T cell can be efficiently and conveniently identified.

Abstract: The invention relates to a method and a device for the automatic processing of biological samples which can have a relatively large volume and which can be further processed by a microfluidic system. A container is provided with a filter and can be or is closed. A biological sample is introduced into the container. An inlet or outlet for liquids is mounted downstream of the filter. In order to carry out processing, liquids that are present in the container are not only sucked off through the filter and passed on via the inlet or outlet, but liquids required for processing are also pumped into the container through the filter. The container can be connected to a microfluidic system in a relatively easy manner since only one conduit is required for an automated processing.

Abstract: The invention generally relates to methods and kits for isolating nucleic acids from an organism. In certain embodiments, methods of the invention involve contacting a plurality of lytic enzymes to an organism, thereby lysing a cell wall of the organism to release the nucleic acid, and introducing at least one agent to separate the nucleic acid from the lysed cells, thereby isolating the nucleic acid.

Abstract: The present invention relates to a method of enriching for membranous microvesicles relative to the cellular population in a biological sample. More particularly, there is provided a method for enriching for exosomes from plasma. In a related aspect, there is provided a method of reducing the concentration of cellular and cellular derived molecules in a biological sample. Still further, the present invention provides methods for selectively isolating mRNA subpopulations from exosomes. Yet further, there are provided methods of amplifying exosome derived RNA. The method of the present invention is useful in a range of applications including, but not limited to, diagnostic, prognostic, therapeutic, research and development applications, to the extent that the enrichment of exosomes is required.

Abstract: The invention provides pipette tip columns and automated methods for the purification of nucleic acids such as plasmids from unclarified cell lysates containing cell debris as well as methods for making and using such columns. The columns typically include a bed of medium positioned in the pipette tip column, above a bottom frit and with an optional top frit.

Abstract: Disclosed is the use of a separation agent including fibril cellulose for phase separation, as well as to a method for the separation of at least one liquid phase from at least one phase selected from a second liquid phase, solid phase and semi-solid phase, where the method includes the steps of incorporating a separation agent including fibril cellulose to a mixture where separation of the phases is desired, followed by formation of phases and removing the phases.

Abstract: Removal of genomic DNA from biological samples using cation exchange is described.

Abstract: The present invention provides a method for isolating nucleic acids from a veterinary whole blood sample, said method comprising at least the following steps a) preparing a binding mixture comprising—the lysed sample—at least one chaotropic agent—at least one alcohol—at least one polyoxyethylene fatty alcohol ether; b) passing the binding mixture through a column comprising a nucleic acid binding solid phase thereby binding the nucleic acids to the nucleic acid binding solid phase; c) optionally washing the nucleic acids while being bound to the solid phase; d) optionally eluting the nucleic acids from the solid phase. It was found that the addition of the specific non-ionic detergent overcomes the problems of the prior art methods, wherein the column clogs what prevents the efficient nucleic acid isolation from this difficult sample.

Abstract: This invention relates to a method, kit and system for collecting and processing of samples to release and treat DNA and RNA for gene sequence detection. The method described here in provides for rapid and convenient release, and recovery of DNA and RNA from tissues and cellular materials.

Abstract: A process for the extraction of pDNA from cells is provided. In one aspect, the process comprises heating a liquid comprising the cells to an average temperature of from 95° C. to about 120° C. for a time of less than 10 seconds. In certain preferred aspects, the pDNA is extracted by the use of flow-through apparatus.

Abstract: A reagent for oligonucleotide synthesis or purification, wherein the reagent has a structure of: X—C—L—H??(Formula A) wherein X is a phosphoramidite group, an H-phosphonate group, an acetal group, or an isocyanate; C is a direct bond or a cleavable adaptor represented by —Ca—Cb—; L is a hydrocarbyl chain; and H is a terminal alkyne or an activated cyclooctyne. The reagent of Formula (A) can be used in the synthesis and purification of oligonucleotides.

Abstract: There is provided a nucleic acid extraction method applicable to microbes in a relatively wide range, and capable of rapidly extracting nucleic acid. The nucleic acid extraction method comprises the steps of introducing a cell suspension into a vessel, sealing the vessel, and preheating a heater up to a set temperature not lower than 100° C. Further, the method comprises the step of bringing the vessel into contact with the heater heated up to the set temperature, thereby heating the cell suspension housed in the vessel up to a prescribed highest temperature at not lower than 100° C. with the vessel held in a sealed state.

Abstract: A coating of a random copolymer of acrylamide and a second monomer, e.g. glycidoxylmethacrylate, for a silica surface is described. The coating is applied to chromatographic support structures having silica based surfaces. The coating is functionalized to produce protein chromatography matrices that are particularly useful for extracting trace amounts of biomarker molecules from biological samples.

Abstract: The invention relates to methods for isolating and/or identifying nucleic acids. The invention also provides kits for isolating and/or identifying nucleic acids.

Abstract: The present invention relates to a method of ensuring the effectiveness of the extraction workup from a biological sample of nucleic acid. The inventive method is able to distinguish between possible defects in the extraction of nucleic acid from a sample and possible defects in a subsequent amplification step. The present invention also relates to a packaged array for extracting nucleic acid from a biological sample.

Abstract: A protein-immobilizing solid phase is a protein-immobilizing solid phase comprising an mRNA-nucleic acid linker-protein complex, obtained by linking the mRNA and the protein encoded by that mRNA through the nucleic acid linker, immobilized on the solid phase, wherein the nucleic acid linker has a photocleavage site and a solid phase binding site.

Abstract: A process for separating organic compounds from a mixture by reverse-phase displacement chromatography, including providing a hydrophobic stationary phase; applying to the hydrophobic stationary phase a mixture comprising organic compounds to be separated; displacing the organic compounds from the hydrophobic stationary phase by applying thereto an aqueous composition comprising a non-surface active hydrophobic anionic displacer molecule and about 10 wt % or less of an organic solvent; and collecting a plurality of fractions eluted from the hydrophobic stationary phase containing the separated organic compounds; in which the non-surface active hydrophobic anionic displacer molecule comprises a hydrophobic anion and a counterion, CI, having the general formula A or B, as defined in the disclosure: [CM][Cl]d [CM-R*—CM?][Cl]d A B.

Abstract: The invention relates to a method for filtering nucleic acids, to a kit for carrying out the method according to the invention and to a novel use of magnetic particles for filtering a biological sample. The method according to the invention comprises the following steps: a) the sample is held in an aqueous solution; b) lysing of the sample; c) separation of cellular debris; and d) the nucleic acids are isolated from the solution, steps (a) to (c) taking place under non-chaotropic conditions.

Abstract: The invention relates to a nucleic acid preparation with a content of below 1% protein, preferably below 0.1% protein, free of ethidium bromide, phenol, cesium chloride and detergents based on octyl phenol polyethylene glycol ether)n and with a content of below 1 EU/mg DNA of endotoxins. Said preparation is suitable as a drug particularly in gene therapy.

Abstract: The present invention relates to a simple and efficient method to isolate and purify nucleic acids, preferably genomic DNA, from complex samples compared with available methods, by using a ligand which relies on hydrogen bonding to purify the nucleic acids. Preferably the ligand is bound to magnetic beads/particles. More closely the method comprises adding a sample comprising nucleic acid to a polymer having neutral charge; reversibly binding said nucleic acid to said polymer by hydrogen bonding under pH conditions <5; washing said polymer; and eluting said nucleic acid from said polymer under conditions of pH >5. The method is very suitable for sample preparation of nucleic acids, for example for PCR applications.

Abstract: Sample preparation processes for in situ RNA or DNA analysis, methods and compositions therefor are provided. Processes provided herein allow DNA or RNA analysis to be carried out in the same tube or on an aliquot of the prepared sample without centrifugation or extraction. The preparation process can be carried out at room temperature in as little as seven minutes and is amenable to high throughput processing using manual or robotic platforms.

Abstract: Removable cartridges are used on automated flow-through systems for the purpose of extracting and purifying genetic material from complex matrices. Different types of cartridges are paired with specific automated protocols to concentrate, extract, and purifying pathogenic or human genetic material. Their flow-through nature allows large quantities sample to be processed. Matrices may be filtered using size exclusion and/or affinity filters to concentrate the pathogen of interest. Lysed material is ultimately passed through a filter to remove the insoluble material before the soluble genetic material is delivered past a silica-like membrane that binds the genetic material, where it is washed, dried, and eluted. Cartridges are inserted into the housing areas of flow-through automated instruments, which are equipped with sensors to ensure proper placement and usage of the cartridges. Properly inserted cartridges create fluid- and air-tight seals with the flow lines of an automated instrument.

Abstract: An apparatus and a method for obtaining a (poly)nucleotide sequence of interest include steps of cultivating hosts cells to produce a nucleotide sequence of interest and harvesting these cells, introducing these cells in a passageway and disintegrating them in a continuous process. In the continuous process, performing in the passageway a precipitation of contaminants by a mixing of the disintegrated cells with a solution containing one or more salt(s) and obtaining a mixture and allowing a precipitate to separate from the solution of this mixture, preferably to float and/or to sediment from the solution of this mixture for 1-48 hours and pumping out a soluble material from this solution, while excluding recovering the precipitate.

Abstract: The present invention pertains to a method for isolating nucleic acids by size from a sample comprising nucleic acids of different sizes using an anion exchange matrix, wherein nucleic acids of a preselected size or a preselected size range are isolated by varying the pH value during elution and/or binding.

Abstract: The identification and evaluation of mRNA and protein targets associated with mRNP complexes and implicated in the expression of proteins involved in common physiological pathways is described. Effective targets are useful for treating a disease, condition or disorder associated with the physiological pathway.

Abstract: The present invention solves the problem of isolating nucleic acids from cells in the presence of the growth medium. The invention is particularly useful for isolating extrachromosomal replicons such as plasmids. Cells are lysed in the presence of the medium in which they were grown and nucleic acids are isolated using a pipette tip column. A liquid handling robot can be used to isolate nucleic acids from multiple samples simultaneously without the need for human intervention.

Abstract: The present invention relates to a method for purifying a defined amount of nucleic acids from a nucleic acid-containing sample, which has at least the following steps: (a.) contacting the nucleic acid-containing sample with a defined amount of a nucleic acid binding phase with the following features: (i) the nucleic acid binding phase has nucleic acid binding ligands that have at least one protonatable group; (ii) the nucleic acid binding ligands are bound to a carrier; (iii) the nucleic acid binding phase has a surface with a low charge density, wherein the amount of nucleic acids in the sample exceeds the binding capacity of the amount of nucleic acid binding phase used; (b.) binding of the nucleic acids to the nucleic acid binding phase at a pH (binding pH) that is below the pKs value of at least one of the protonatable groups; (c.) elution of the nucleic acids at a pH that is above the binding pH, wherein a defined amount of nucleic acids is obtained.

Abstract: In one aspect of the invention, methods for analyzing nucleic acid sample are provided. In a preferred embodiment, nucleic acids are selected using affinity matrices prior hybridization with a microarray.

Abstract: A polyelectrolyte-coated particle, devices for using the particle, methods for using the particle for separating PCR reaction products and/or DNA sequencing reaction products, and compositions for coating the particle are provided.

Abstract: The present invention provides methods, compositions, and kits for generating rRNA-depleted samples and for isolating rRNA from samples. In particular, the present invention provides compositions comprising affinity-tagged antisense rRNA molecules corresponding to substantially all of at least one rRNA molecule (e.g., 28S, 26S, 25S, 18S, 5.8S and 5S eukaryotic cytoplasmic rRNA molecules, 12S and 16S eukaryotic mitochondrial rRNA molecules, and 23S, 16S and 5S prokaryotic rRNA molecules) and methods for using such compositions to generate rRNA-depleted samples or to isolate rRNA molecules from samples.

Abstract: Methods for isolating nucleic acids using a solid phase having alkylene sulfonyl-containing compounds on its surface are provided. In one embodiment, the method comprises: contacting a sample containing nucleic acids with a solid phase having alkylene sulfonyl-containing compounds on its surface in a first aqueous solution to provide a loaded solid phase; washing the loaded solid phase with a second aqueous solution to provide a washed solid phase; and eluting the washed solid phase with a low ionic strength liquid to obtain the isolated nucleic acids. Kits containing a solid phase having alkylene sulfonyl-containing compounds also are provided.

Abstract: Provided herein is technology relating to isolating nucleic acids. In particular, the technology relates to methods and kits for serial extraction of multiple DNA targets from a human stool sample.

Abstract: The present invention pertains to a method for isolating extracellular nucleic acids from a sample, wherein said sample is optionally stabilized, by binding the extracellular nucleic acids to a solid phase which carries anion exchange groups, comprising the following steps: binding the extracellular nucleic acids to the solid phase in a binding mixture having a first pH which allows binding the extracellular nucleic acids to the anion exchange groups of the solid phase; wherein the sample makes up at least 85% of the volume of the binding mixture; separating the solid phase with the bound extracellular nucleic acids; optionally washing the extracellular nucleic acids; optionally eluting extracellular nucleic acids from the solid phase. The method has the advantage that large sample volumes can be processed and that extracellular nucleic acids can be isolated rapidly with a high yield. The method is particularly suitable for automatable processes.

Abstract: The present invention relates to matrix materials suitable for use in purifying and/or isolating nucleic acids from a biological sample, which matrix comprises a surface comprising at least one element selected from the group consisting of Germanium, Tin and/or Lead, or at least one salt thereof, and methods related therewith.

Abstract: The present invention is directed to a method for separation of an oligonucleotide from a mixture using a biphasic mobile phase/stationary phase liquid-liquid chromatography system. A first mobile phase contains the oligonucleotide and the stationary phase contains an exchanger substance that removably binds to the target oligonucleotide. The mobile phase is caused to flow in contact with the stationary phase in a liquid-liquid chromatography apparatus such that the oligonucleotide becomes bound to the exchanger substance in the liquid stationary phase. The oligonucleotide is then displaced from the liquid stationary phase into a second liquid mobile phase by means of a displacer substance able to displace the oligonucleotide from the stationary phase into the second mobile phase.

Abstract: A method is provided for purifying nucleic acid from a sample in a microfluidic device. The method can be used to purify nucleic acids from any source known in the art that comprises nucleic acids, such as prokaryotic or eukaryotic organisms, viruses, cell, tissues, organs, etc. In a specific example, the tissue is whole blood. The method for purifying nucleic acid may run fully automated in the microfluidic device.

Abstract: The invention generally provides a method for the sample preparation for a subsequent preparation, processing or analysis method of a sample containing an at least one species of nucleic acid and/or one species of protein, whereby the method comprises the following steps: A) providing a sample which contains at least one species of a nucleic acid and/or of a protein, B) contacting the sample with a fluid or solid composition to produce a fluid sample preparation, whereby the composition contains at least a nitrogenous compound, which is selected from the group consisting of a) polyamines, b) amino acids, and oligo- and polypeptides, c) nitrogenous heterocyclic compounds, including homo-oder heteropolymeres, which comprise these nitrogenous compounds, d) amines of the type R1R2NR3, whereby R1, R2 and R3 are chosen independently from one another from the group consisting of H, C1-C5-alkyl groups and aryl groups, whereby R1, R2 and R3 are not H simultaneously, e) carbonxylic acid amides, f) inorganic ammonium sa

Abstract: The present disclosure relates to a method and apparatus for separating nucleic acids. A carrier may include a porous microbead having cation-exchangeable groups attached to the surface of the porous microbead. Capturing chains modified with positively charged functional groups and having a base sequence complementary to a target nucleic acid chain sequence are immobilized on to the surface of the porous microbead. In various embodiments, capturing chains are immobilized on to the surface of the porous microbead through an ion exchange bond or a covalent bond with the cation-exchangeable groups of the porous microbead. In some cases, the porous microbead has a number of through pores adapted to permit a solution to pass rapidly through the through pores and a number of diffusive pores adapted to permit a solute of the solution to diffuse into the diffusive pores.

Abstract: Methods of using hypermethylated transposons to create genetically modified animals that express interfering RNAs are described.

Abstract: The invention provides methods for isolating cell-type specific mRNAs by selectively isolating ribosomes or proteins that bind mRNA in a cell type specific manner, and, thereby, the mRNA hound to the ribosomes or proteins that bind mRNA. Ribosomes, which are riboprotein complexes, bind mRNA that is being actively translated in cells. According to the methods of the invention, cells are engineered to express a molecularly tagged ribosomal protein or protein that binds mRNA by introducing into the cell a nucleic acid comprising a nucleotide sequence encoding a ribosomal protein or protein that binds mRNA fused to a nucleotide sequence encoding a peptide tag. The tagged ribosome or mRNA binding protein can then be isolated, along with the mRNA bound to the tagged ribosome or mRNA binding protein, and the mRNA isolated and further used for gene expression analysis.

Abstract: The present invention provides a chromatography column and a method for isolating nucleic acid molecules. In one embodiment, the present invention provides a double-layer column of a first anion exchange membrane and a second serially coupled silica membrane. Upon flowing a nucleic acid-containing solution through the first anion exchange membrane, the nucleic acid binds to and then elutes from the first membrane. The eluted solution then flows serially through the second silica membrane, which the nucleic acid binds to and then elutes from. Due to this novel serial coupled double-layer principle, the present invention is particularly suitable for co-isolating RNA and DNA, for isolating nucleic acid embraced by proteins, e.g., viruses, and for isolating diluted nucleic acid in a large volume, e.g., plasma. In addition, the eluted nucleic acid is ready for downstream applications.

Abstract: A method for preparing a stool sample without any need for complicated operations is provided which is capable of efficiently recovering a nucleic acid originating from mammalian cells, such as the cells exfoliated from the large intestine, in the stool. A solution for preparing a stool sample and a kit for stool collection are also provided. The collected stool is mixed with a solution for preparing a stool sample which has a water-soluble organic solvent as an active ingredient. A method is disclosed for recovering a nucleic acid including recovering a nucleic acid originating from indigenous enteric bacterium and a nucleic acid originating from an organism other than indigenous enteric bacterium at the same time from the stool sample prepared by the preparation method.

Abstract: The present invention is directed to variant squalene synthase enzymes, including Saccharomyces cerevisiae squalene synthase enzymes, and to nucleic acid molecules encoding these variant enzymes. These variant enzymes produce squalene at a lower rate than the wild-type enzyme, allowing more farnesyl pyrophosphate to be utilized for production of isoprenoid compounds, while still producing sufficient squalene to allow the S. cerevisiae cells to grow without the requirement for supplementation by sterols such as ergosterol. These variant enzymes, therefore, are highly suitable for the efficient production of isoprenoids.

Abstract: Described herein are techniques for assembling a polynucleotide encoding a transcription activator-like effector nucleases (TALEN). The techniques ligate and digest necessary modules for a TALEN assembly in one reactor or system. Methods and Kits for generating a TALEN are also described.

Abstract: A method for nucleic acid isolation comprising: receiving a binding moiety solution within a process chamber; mixing the binding moiety solution with a biological sample, within the process chamber, in order to produce a moiety-sample mixture; incubating the moiety-sample mixture during a time window, thereby producing a solution comprising a set of moiety-bound nucleic acid particles and a waste volume; separating the set of moiety-bound nucleic acid particles from the waste volume; washing the set of moiety-bound nucleic acid particles; and releasing a nucleic acid sample from the set of moiety-bound nucleic acid particles. The method preferably utilizes a binding moiety comprising at least one of poly(allylamine) and polypropylenimine tetramine dendrimer, both of which reversibly bind and unbind to nucleic acids based upon environmental pH.

Abstract: The invention provides methods of preparing nucleic acids, such as RNA molecules, of a defined size or range of sizes. The invention provides compositions, methods and kits for use in the production and preparation of small RNA molecules (including without limitation micro-RNA, siRNA, d-siRNA and e-siRNA) and other nucleic acids of various sizes.

Abstract: A method for isolating polynucleotides, such as DNA, RNA and hybrids thereof from an aqueous solution containing polynucleotides by reversibly binding the polynucleotides to silica-coated magnetic particles in the presence of a salt and non-ionic hydratable additive is disclosed. The salt and non-ionic hydratable additive concentrations are adjusted to levels that result in adhesion of the nucleic acid to the particles without degradation or precipitation of the nucleic acid.

Abstract: The present invention relates to a device for rapid isolation of target molecules from cell lysates and other liquid mixtures comprising particulate material, a method for isolating the target molecules, in particular nucleic acids, using said device and a kit for carrying out said method comprising said device.