Abstract: The present invention is directed to methods for increasing the efficiencies with which recombinant adeno-associated virus (rAAV) are packaged, so as to increase their production titers. More specifically, the invention relates to a method for increasing the production titer of rAAV by transfected cells by increasing the ionic strength of the cell culture media through the administration of additional ions.

Abstract: In a liquid stirring method, after a second liquid is discharged into a reaction container accommodating a first liquid from a dispensing probe provided with a dispensing tip at the leading end thereof, a mixture of the first liquid and second liquid in the container is stirred by being sucked out and discharged by the dispensing probe. The number of stirrings through sucking out and discharging is changed according to the total volume of the first liquid and second liquid. If the total volume of the first liquid and second liquid is below a preset threshold, sucking out and discharging is repeated for a prescribed number of times.

Abstract: A microscale sampling device including a frame is provided in the present invention, a sample container, a communicating channel and a resistance channel are defined in the frame. At least one sampling chamber is defined in the communicating channel. An end of the communicating channel is communicated with the sample container and the communicating channel is arranged below the sample container. An end of the resistance channel is communicated with the sampling chamber, and the other end of the resistance channel is communicated to an output joint. The resistance channel is shaped with at least one discontinuous shape change.

Abstract: Aspects of the disclosure include methods for the preparation of a polynucleotide. In some embodiments, the method includes contacting a first polynucleotide composition including: a polynucleotide having a sequence of 7 or more nucleoside subunits and at least two of the nucleoside subunits are joined by a N3??P5? thiophosphoramidate inter-subunit linkage; and non-target synthetic products and reagents; with a multivalent cation salt to precipitate a polynucleotide salt including at least one multivalent cation counterion; and separating the polynucleotide salt from the contacted first polynucleotide composition to produce a second polynucleotide composition including the polynucleotide salt. In certain embodiments, the method further includes contacting the polynucleotide salt with a reverse phase chromatography support; and eluting from the chromatography support a third polynucleotide composition including the polynucleotide.

Abstract: The present invention relates to a method for selectively depleting animal nucleic acids from non-animal nucleic acids in a sample, which comprises animal cells and at least one further type of cells, selected from microbial cells and plant cells or a combination thereof, to a lysis solution A to be used in and to a kit to carry out said method as well as to the use of a particular silica membrane as both, a filtration matrix for separating essentially intact microbial and/or plant cells from lysed animal cells and an adsorption matrix for nucleic acids, in particular in a method according to the present invention.

Abstract: Methods for processing polynucleotide-containing biological samples, and materials for capturing polynucleotide molecules such as RNA and/or DNA from such samples are provided. The RNA and/or DNA is captured by polyamindoamine (PAMAM (Generation 0)) bound to a surface, such as the surface of magnetic particles. The methods and materials have high efficiency of binding RNA and of DNA, and of release, and thereby permit quantitative determinations. The binding particles can be coated with PAMAM (Generation 0) dendrimer which is amide-bonded to the binding particles, wherein at least a portion of the nucleic acids in the biological sample become reversibly bonded to the binding particles.

Abstract: A method of recovering a bead support from an emulsion includes supplying an aqueous surfactant solution into a centrifuge tube; supplying a hydrophobic liquid over the surfactant solution in the centrifuge tube, wherein a ratio of the volume of the aqueous surfactant solution to the volume of the hydrophobic liquid is not greater than 0.5; and applying an emulsion over the hydrophobic liquid while centrifuging, the emulsion comprising a dispersed aqueous phase including the bead support, the emulsion breaking and material of the dispersed phase preferentially partitioning to the surfactant solution.

Abstract: The present invention is directed to fluorescent molecular sensor based on Thiazole Orange for protein detection. Interaction of the protein target with the molecular sensors of this invention results in a significant increase in the fluorescence emission. The generation of light output signal enables one to detect protein biomarkers associated with different diseases or detecting the protein of interest also in living cells.

Abstract: A gold nanoparticle-based assay for the detection of a target molecule, such as Hepatitis C Virus (HCV) RNA in serum samples, that uses positively charged gold nanoparticles (AuNPs) in solution based format. The assay has been tested on 74 serum clinical samples suspected of containing HCV RNA, with 48 and 38 positive and negative samples respectively. The developed assay has a specificity and sensitivity of 96.5% and 92.6% respectively. The results obtained were confirmed by Real-Time PCR, and a concordance of 100% for the negative samples and 89% for the positive samples has been obtained between the Real-Time PCR and the developed AuNPs based assay. Also, a purification method for the HCV RNA has been developed using HCV RNA specific probe conjugated to homemade silica nanoparticles. These silica nanoparticles have been synthesized by modified Stober method. This purification method enhanced the specificity of the developed AuNPs assay.

Abstract: Methods and systems for processing polynucleotides (e.g., DNA) are disclosed. A processing region includes one or more surfaces (e.g., particle surfaces) modified with ligands that retain polynucleotides under a first set of conditions (e.g., temperature and pH) and release the polynucleotides under a second set of conditions (e.g., higher temperature and/or more basic pH). The processing region can be used to, for example, concentrate polynucleotides of a sample and/or separate inhibitors of amplification reactions from the polynucleotides. Microfluidic devices with a processing region are disclosed.

Abstract: Disclosed is a process for purifying nucleic acids, especially plasmid DNA, from a nucleic acid-containing, biologic sample, consisting of (a) preparation of a fluid sample containing nucleic acid and endotoxin each in dissolved form; (b) precipitation at least of nucleic acid and endotoxin from the fluid sample; (c) washing of the components of the fluid sample precipitated out in step b) in order to at least partially remove the endotoxin with at least one washing solution, which contains at least one amine compound with at least two carbon atoms and with a molar mass of ?500 g/mol and; at least one organic solvent different from the aforementioned amine compound and has a pH-value (20° C.) in the range from pH 3.0 to pH 8.5, and; (d) dissolution of the remaining nucleic acid from the washed, precipitated constituents from step c) using a dissolving buffer and collection of the dissolved nucleic acids in a separate receptacle. The process is suitable for the effective and economical removal of endotoxins.

Abstract: The present invention relates to a new method for enriching and/or isolating nucleic acids from microbial cells which comprises filtering a liquid sample through a nucleic acid-binding matrix which has a pore size small enough to retain microbial cells, lysing the microbial cells on the matrix to release the nucleic acids from the microbial cells, binding the nucleic acids to the matrix and subsequently eluting the DNA. The invention also relates to a method for enriching and/or isolating nucleic acids from microbial cells which are present in a liquid sample that comprises microbial cells and higher eukaryotic cells and/or tissues. The invention also provides a cartridge for carrying out the methods of the invention. Finally, the invention relates to kits for carrying out the methods of the invention.

Abstract: The present invention provides a special reagent for the pretreatment of sample materials. The pretreatment with the reagent according to the invention enables the isolation of nucleic acids by a uniform method from very diverse sample materials, in particular different bioprocess samples, even if the nucleic acids are only present in small amounts in the sample materials. The reagent used for the pretreatment comprises, as key component, at least one compound comprising an amino group. The invention further relates to methods of purification of nucleic acids, in which the sample material is pretreated correspondingly, and suitable kits.

Abstract: The present disclosure relates to systems and methods for nucleic acid isolation from cellular samples. In particular, the present disclosure provides systems and methods for lysing cells and recovering nucleic acids.

Abstract: The present invention relates to compositions, kits and methods for making and using RNA compositions comprising in vitro-synthesized ssRNA inducing a biological or biochemical effect in a mammalian cell or organism into which the RNA composition is repeatedly or continuously introduced. In certain embodiments, the invention provides compositions and methods for changing the state of differentiation or phenotype of a human or other vertebrate cell. For example, the present invention provides mRNA and methods for reprogramming cells that exhibit a first differentiated state or phenotype to cells that exhibit a second differentiated state or phenotype, such as to reprogram human somatic cells to pluripotent stem cells.

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: Parallel isolation of a double-stranded nucleic acid and a single-stranded nucleic acid is possible from a sample that contains these acids, without separating the acids, by mixing the sample with a lysis buffer having high salt concentration or low salt concentration, or having a proteolytic enzyme. The sample that contains nucleic acid before its lysis, or the sample that has already been lysed or homogenized, is adjusted with a binding buffer in such a manner that the total nucleic acid is adsorbed onto a solid carrier. The binding buffer contains at least one non-ionic detergent in a high concentration. With the exception of the detergent, the sample contains no other non-acidic organic component miscible in water. The carrier with the adsorbed total nucleic acid is removed. The adsorbed total nucleic acid is washed and eluted.

Abstract: The invention is directed to compositions and methods for rapidly and efficiently extracting nucleic acids and/or targeted nucleic acids sequences from biological samples. The methods of the invention comprise combining the sample with a buffer and magnetic silicon beads and concentrating the beads with a magnet or other electrical field. Liquid may be removed, or not, and an alkaline buffer is added followed by magnetic carboxy beads in a binding buffer so that nucleic acids transfer to the carboxy beads, which can be easily and quickly isolated once again with a magnet. Total nucleic acid extraction is greatly enhanced. Extracted nucleic acids can be analyzed, for example, by PCR wherein the nucleic acids can be identified and characterized. Carboxy beads may also contain a ligand so as to target specific nucleic acid sequences. The invention is also directed to kits comprising the tools and compositions for performing the methods of the invention.

Abstract: The invention is directed to compositions and methods for rapidly and efficiently extracting nucleic acids and/or targeted nucleic acids sequences from biological samples. The methods of the invention comprise combining the sample with a buffer and magnetic silicon beads and concentrating the beads with a magnet or other electrical field. Liquid may be removed, or not, and an alkaline buffer is added followed by magnetic carboxy beads in a binding buffer so that nucleic acids transfer to the carboxy beads, which can be easily and quickly isolated once again with a magnet. Total nucleic acid extraction is greatly enhanced. Extracted nucleic acids can be analyzed, for example, by PCR wherein the nucleic acids can be identified and characterized. Carboxy beads may also contain a ligand so as to target specific nucleic acid sequences. The invention is also directed to kits comprising the tools and compositions for performing the methods of the invention.

Abstract: An apparatus for extracting an analyte from a liquid sample having a container with an entrance, an exit, and a passage therebetween for passage of a liquid sample containing an analyte, the container having a full diameter bed region and a reduced diameter bed region. The container includes a layered construction extending across the passage, having from top to bottom one or more of: (i) an upper flow distributor/support layer, (ii) an upper compression layer, (iii) an extraction layer of microparticulate extraction medium adjacent to the layer (ii), and (iv) a lower compression layer located adjacent to the extraction layer (iii), optionally including one or more air gap layers. At least some of the layers are located in the full diameter bed region, and some of the layers are located in the reduced diameter bed region. The apparatus may have a plurality of containers arranged in an array and/or in series so as to provide multi-stage filtration or extraction.

Abstract: Wound dressing articles comprising a nonwoven web comprising a plurality of fibers having grafted pendant hydrophilic groups, methods that use high energy irradiation for making a plurality of fibers having grafted pendant hydrophilic groups, useful for making wound dressing articles.

Abstract: Methods and compositions are described herein for protecting RNA from autocatalytic and divalent cation induced degradation in an aqueous solution.

Abstract: The invention related to a method for the stabilization, purification or/and isolation of nucleic acids from material samples, in particular, stool samples, which can contain impurities and inhibitors or interfering substances. The invention further relates to a reagent kit for carrying out this method. The basis of the invention is, in particular, a method for purification, stabilization or/and isolation of nucleic acids from material samples, whereby a buffer is added to the sample containing the nucleic acids, with a pH value of 2 to 7, a salt concentration of at least 100 mM, or/and a phenol neutralizing substance. According to the invention, pure nucleic acids which may be amplified can be obtained from faecal samples by a simple method, which are suitable for diagnostic proof of infection, in particular, bacterial or viral infection, or mutation, in particular, for tumor-specific DNA mutations.

Abstract: The present invention relates to polymorphic forms of 5-Azacytidine and the process for preparation thereof. The present invention further relates to Crystalline 5-azacytidine 5 designated as Form-SA-1 characterized by an X-ray powder diffraction pattern having at least four characteristic diffraction angle peaks at about 12.00, 12.60, 13.90, 15.15 and 31.40±0.20 2?°, which is useful as active pharmaceutical ingredient in pharmaceutical compositions for the treatment of myelodysplastic syndrome.

Abstract: A method for processing a nucleic acid, in which the nucleic acid is exposed to an aqueous medium which includes a polyol in sufficient proportion for at least a portion of the nucleic acid to enter or remain in an extra-solution phase. Thus, a polyol may be used to bind a nucleic acid which is in solution to a solid support or to wash a nucleic acid on a solid support while maintaining it on the support. The polyol may for example be a C2-C10 alkanediol.

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 relates to methods and systems for microfluidic DNA sample preparation. More specifically, embodiments of the present invention relate to methods and systems for the isolation of DNA from patient samples on a microfluidic device and use of the DNA for downstream processing, such as performing amplification reactions and thermal melt analysis on the microfluidic device.

Abstract: [Object] To provide a method of purifying nucleic acids where the operation is simple and the nucleic acids can be extracted in a short time with high efficiency. [Solving Means] A method of purifying nucleic acids including the step of adsorbing substances in a sample containing nucleic acids with an ion exchange resin 10 including a positive ion exchange resin and a negative ion exchange resin. As the positive ion exchange resin, a first positive ion exchange resin and a second positive ion exchange resin having an exclusion limit molecular weight lower than that of the first positive ion exchange resin may be used.

Abstract: The present invention relates to a method for the lysis of cells, especially bacterial cells and the isolation of nucleic acids. The sample is treated with lysis solution that comprises at least one liquid that is not miscible with water and heated. Afterwards the mixture is cooled down and water is added so that after cooling a two phase system is generated. The nucleic acids can be found in the aqueous phase.

Abstract: The invention discloses a method of extracting nucleic acid analyte solution from biological sample, comprising: placing the biological sample into a heating container; optionally, adding solvent medium into the biological sample; heating the biological sample under high pressure; optionally, centrifuging the biological sample; and obtaining the solution including the nucleic acid analyte. The invention also discloses a method of detecting the nucleic acid analyte obtained by said extraction method.

Abstract: The present invention pertains to a method of isolating RNA from a sample comprising RNA, and DNA, comprising: a) adding an acidic denaturing composition comprising a chaotropic agent and phenol to the sample; b) adding a water-insoluble organic solvent and separating the resulting phases thereby forming a multi-phase mixture comprising an aqueous phase, optionally an interphase and an organic phase, wherein the RNA is concentrated in said aqueous phase and DNA and proteins are concentrated in said organic phase and/or in said interphase; and c) isolating said RNA from said aqueous phase, wherein at least one cationic detergent is added before separating the phases. It was found that the addition of at least one cationic detergent considerably reduces the amount of DNA in the aqueous, RNA containing phase. Therefore, the present invention allows to easily isolate pure RNA which comprises considerably less DNA contaminations.

Abstract: The invention relates to a chromatographic device for isolating and/or purifying double-stranded nucleic acids, preferably double-stranded DNA, from a mixture of such nucleic acids with single-stranded nucleic acids, oligonucleotides, mononucleotides, salts and/or other such impurities. The invention also relates to a method for chromatographically isolating and/or purifying same, and to a kit for this purpose.

Abstract: It is an object of the present invention to provide a method for eluting an adsorbed protein that suppresses a decrease in protein adsorption ability, in a method for purifying a protein using a protein-adsorbing porous membrane. The present invention provides a method for purifying a protein, comprising an adsorption step and an elution step, wherein in the elution step, at least one eluent is passed in the opposite direction with respect to the direction of the passage of a stock solution containing an adsorption target protein, in the adsorption step.

Abstract: A cDNA synthesis method includes: mixing a lysis solution containing a chaotropic substance and a nucleic acid-binding solid-phase carrier in a sample containing a ribonucleic acid (RNA), thereby adsorbing the RNA on the carrier; reverse-transcribing the RNA adsorbed on the carrier while keeping the RNA adsorbed on the carrier in a reverse transcription reaction mixture, thereby synthesizing cDNA; and eluting the synthesized cDNA with an eluent.

Abstract: The present invention is directed to methods to prepare a DNA molecule or a plurality of DNA molecules by random fragmentation. In some embodiments, the present invention regards preparing a template for DNA sequencing by random fragmentation. In specific embodiments, the random fragmentation comprises chemical fragmentation, mechanical fragmentation, or enzymatic fragmentation. In further specific embodiments, a universal sequence is attached to the 3? end of the DNA fragments, such as by ligation of an adaptor sequence or by homopolymeric tailing with terminal deoxynucleotidyltransferase. In other embodiments, a library is prepared with methods of the present invention.

Abstract: The present disclosure relates to systems and methods for nucleic acid isolation from cellular samples. In particular, the present disclosure provides systems and methods for lysing cells and recovering nucleic acids.

Abstract: Composition based on polynucleotides extracted from natural sources for use in therapeutic treatment and/or as a therapeutic co-adjuvant in the treatment of degenerative diseases of the joints, in particular osteoarthritis.

Abstract: A method for i) parallel isolation of a double-stranded and/or a single-stranded nucleic acid and/or ii) selective removal of a double-stranded nucleic acid from a mixture of a double-stranded and a single-stranded nucleic acid or from a source comprising a double-stranded and a single-stranded nucleic acid includes absorbing the double-stranded nucleic acid onto a first solid carrier, while the single-stranded nucleic acid is not adsorbed and remains in solution, removing the first carrier with the adsorbed nucleic acid from the solution, mixing the solution comprising the single-stranded nucleic acid with an alcoholic solution having a concentration of 1 to 90 vol.-%, and contacting the resulting solution with second solid carrier, to absorb the single-stranded nucleic acid onto the second solid carrier.

Abstract: Disclosed are methods for isolating polynucleotides (e.g., RNA and DNA) from a cell-containing sample. The disclosed methods employ polyamidoamine (PAMAM) dendrimer molecules bound to the surface of binding particles to capture and isolate the polynucleotides from the samples. The polynucleotides are released from the binding particles by contacting the binding particles with a release solution having a basic pH.

Abstract: The present invention provides methods, compositions, kits, systems and apparatus that are useful for isolating nucleic acid molecules from a sample. In particular, the methods generally relate to normalizing the concentration of target nucleic acid molecules from a sample. In one aspect, the invention relates to purifying a primer extension product from a primer extension reaction mixture. In some aspects, nucleic acid molecules obtained using the disclosed methods, kits, systems and apparatuses can be used in various downstream processes including nucleic acid sequencing.

Abstract: Methods and composition for nucleic acid isolation are provided. In one embodiment, the invention provides a method for nucleic acid purification from biological samples extracted with phenol-based denaturing solvents, which does not require phase separation or nucleic acid precipitation. Methods according to the invention may also be used for differential isolation of RNA and DNA.

Abstract: This disclosure is directed to an apparatus, system and method for retrieving a target material from a suspension. A system includes a plurality of processing vessels and a collector. The collector funnels portions of the target material from the suspension through a cannula and into the processing vessels. Sequential density fractionation is the division of a sample into fractions or of a fraction of a sample into sub-fractions by a step-wise or sequential process, such that each step or sequence results in the collection or separation of a different fraction or sub-fraction from the preceding and successive steps or sequences. In other words, sequential density fractionation provides individual sub-populations of a population or individual sub-sub-populations of a sub-population of a population through a series of steps.

Abstract: Embodiments of the present disclosure include methods and compositions for functionalizing molecules, such as oligonucleotides, with functional groups, including polyhistidine tags useful in affinity methods. Some embodiments include methods for modifying and purifying complex mixtures of molecules by exchange of functional tags.

Abstract: A method of recovering a bead support from an emulsion includes supplying an aqueous surfactant solution into a centrifuge tube; supplying a hydrophobic liquid over the surfactant solution in the centrifuge tube, wherein a ratio of the volume of the aqueous surfactant solution to the volume of the hydrophobic liquid is not greater than 0.5; and applying an emulsion over the hydrophobic liquid while centrifuging, the emulsion comprising a dispersed aqueous phase including the bead support, the emulsion breaking and material of the dispersed phase preferentially partitioning to the surfactant solution.

Abstract: The invention discloses a method of separating a biomolecule from at least one other component in a liquid, comprising a step of contacting said liquid with a separation matrix comprising a solid support and polymer chains bound to said solid support. The polymer chains comprise units derived from a first monomer of structure CH2?CH-L-X, where L is a covalent bond or an alkyl ether or hydroxysubstituted alkyl ether chain comprising 2-6 carbon atoms, and X is a sulfonate or phosphonate group.

Abstract: Disclosed is a process for retrieval of nucleotide sequence. The process includes mixing iron chloride tetrahydrate with iron (III) chloride hexahydrate in solution; adding ammonium hydroxide to the mixture and stirring to form maghemite nanoparticles; stirring the maghemite nanoparticles in a solution with an inorganic acid, a surfactant and a monomer precursor of a conducting polymer; initiating polymerization of the monomer by adding the inorganic acid and an oxidizing agent to the stirred solution and further stirring to yield Polyaniline/maghemite nanocomposites; adding the nanocomposites to an first aqueous solution of the nucleotide sequence and stirring so as to electrostatically interact the nanocomposites with the nucleotide sequence; and weakening the electrostatic interaction between the nanocomposite and the nucleotide sequence to recover the nanocomposite independently of the nucleotide sequence.

Abstract: Disclosed herein are processes for collecting nucleic acids from particulate samples. One embodiment disclosed herein relates to the use of ultrasonic energy to simultaneously shear large nucleic acid molecules and large particulates to very small sizes prior to or during a chemical binding step to a nucleic acid binding surface. Another embodiment involves crushing the nucleic acid binding surface prior to eluting the bound nucleic acid molecules to enable better wetting of the nucleic acid binding surface and easier diffusion of bound nucleic acid molecules out of the nucleic acid binding surface.

Abstract: Procedure for the specific isolation of total DNA content of bacterial germs of different samples, in the course of which the cells are lysated, the DNA content of the lysate is bound selectively, it is washed and then the desalinated linear polymer nucleic acid is eluted from the binding surface in an aqueous solution. Before cell lysis the nonviable bacterial cells are separated from the viable cells on the basis of their different cell surface physical-chemical characteristics, the viable cells of the sample are kept and then lysated using a mechanical and/or enzymatic, favorably lysozyme enzymatic method. After this exclusively double-stranded DNA deriving from the lysate of viable cells is bound on a —SiO2—TiO2- matrix containing chemically activated —OH and dodecylamine groups, and after washing it, the desalinated linear polymer nucleic acid is eluted in an aqueous solution.

Abstract: Methods and systems for processing polynucleotides (e.g., DNA) are disclosed. A processing region includes one or more surfaces (e.g., particle surfaces) modified with ligands that retain polynucleotides under a first set of conditions (e.g., temperature and pH) and release the polynucleotides under a second set of conditions (e.g., higher temperature and/or more basic pH). The processing region can be used to, for example, concentrate polynucleotides of a sample and/or separate inhibitors of amplification reactions from the polynucleotides. Microfluidic devices with a processing region are disclosed.

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.