Abstract: Disclosed are a method for DNA extraction in a sample for next generation sequencing (NGS) and a method of constructing a NGS library using the extracted DNA. The method for DNA extraction includes: preparing a mixture by mixing a biological sample with a buffer; applying microwaves to the mixture; and recovering DNA. The method of constructing a NGS library includes: extracting DNA according to the method for DNA extraction; amplifying a target DNA using primers; and purifying the amplified product and subjecting the purified product to library pooling.

Abstract: Disclosed are rapid and reproducible methods for separating and purifying nucleic acids from paraffin-containing tissue samples. The disclosed methods involve a melting step, where paraffin-containing tissue samples are heated in the presence of a detergent, causing the paraffin to melt and tissue sample cells to lyse. From the resulting two-phase mixture (i.e., a paraffin phase and an aqueous phase), the aqueous phase is collected for purification of nucleic acid. Protease(s) can be used at one or more points in the separation process to facilitate tissue cell lysis and/or degrade proteins that could degrade nucleic acid or interfere with subsequent genetic manipulation or analysis.

Abstract: The present invention provides methods for the rapid and efficient isolation of small RNA from a biological sample. In particular, small RNA is separated and isolated from large RNA, DNA, proteins, and other macromolecules in the biological sample.

Abstract: The present disclosure generally relates to dry solid matrices for the extraction, stabilization, and storage of nucleic acids, particularly RNA, in a dry format under ambient conditions for a prolonged period of time. Methods for collecting and recovering the nucleic acids stored in the dry solid matrix are also described.

Abstract: A solid matrix for the extraction, stabilization, and storage of nucleic acids is provided. At least one protein denaturant, and at least one acid or acid-titrated buffer reagent are impregnated in a dry state therein the matrix; and the matrix is configured to provide an acidic pH on hydration. The matrix is configured to extract nucleic acids from a sample and stabilize the extracted nucleic acids, particularly RNA, in a dry format under ambient conditions for a prolonged period of time. Methods for collecting and recovering the nucleic acids stored in the dry solid matrix are also described.

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: A method is provided for extraction of chemical compounds from an organism having a cell wall that includes adding nanomaterials, which may be at least one of metallic nanofibers, silica nanofibers, carbon nanofibers and metal doped silica and carbon nanofibers, wherein the nanofibers are generally doped with silver but may also include non-metallic nanofibers doped with metals other than silver, to the organism.

Abstract: A solid substrate for biological sample storage under dry-state and elution of biomolecules is provided. The dry, solid substrate is coated with saccharides, such as monosaccharides, oligosaccharides, polysaccharides or combinations thereof, and the substrate is comprised of one or more protein denaturing agents impregnated therein under a substantially dry state. A method for elution of biomolecules from biological samples is also provided. The compositions disclosed herein provide for enhanced elution and recovery of biomolecules, such as nucleic acids, from the sample. The sample is disposed on a substrate, dried to a substantially dry state; eluted from the biological sample dried on the substrate by rehydrating the substrate in an elution buffer.

Abstract: The present disclosure generally relates to dry solid matrices for the extraction, stabilization, and storage of nucleic acids, particularly RNA, in a dry format under ambient conditions for a prolonged period of time. Methods for collecting and recovering the nucleic acids stored in the dry solid matrix are also described.

Abstract: A solid matrix for the extraction, stabilization, and storage of nucleic acids is provided. At least one protein denaturant, and at least one acid or acid-titrated buffer reagent are impregnated in a dry state therein the matrix; and the matrix is configured to provide an acidic pH on hydration. The matrix is configured to extract nucleic acids from a sample and stabilize the extracted nucleic acids, particularly RNA, in a dry format under ambient conditions for a prolonged period of time. Methods for collecting and recovering the nucleic acids stored in the dry solid matrix are also described.

Abstract: The method for isolating nucleic acids from a food sample comprising the following steps: a) obtaining a food enrichment culture; b) transferring a portion of the food enrichment culture into a reaction vessel thereby providing a food enrichment sample and providing a water-immiscible phase in contact with the food enrichment culture; c) lysing the food enrichment sample to provide a lysed sample; d) isolating nucleic acids from the lysed sample. Food enrichment culture samples are known to contain high concentrations of organic and/or liposoluble inhibitors. By contacting the enrichment culture sample with a water-immiscible phase before the actual DNA extraction procedure starts, part of the lipophilic inhibitors is expected to cross the phase interface due to an enhanced solubility in the organic phase and thereby become depleted.

Abstract: A method of separating nucleic acids from cells, the method comprising incubating a sample comprising cells with a solid substrate that binds to the cells, whereby the cells adhere to the solid substrate; suspending the solid substrate adhered to the cells in a lysis composition comprising about 100 mM to about 300 mM of alkaline metal salt, and having a pH of about 6 to about 8; lysing the cells in the lysis composition to obtain a lysed solution; and obtaining the nucleic acids from the lysed solution; as well as related compositions and kits.

Abstract: Methods are disclosed for isolating nucleic acids from formalin-fixed paraffin embedded (FFPE) tissue samples. Each of tissue samples contains paraffin and a target biological tissue or material, and the method includes the steps of: adding a first reagent and a second reagent to the FFPE tissue sample, the first reagent dissolving the paraffin material and the second reagent lysing the biological tissue; mixing the first reagent, the second reagent, and the FFPE tissue sample to form a first mixture; (2) heating the first mixture at 50-80° C. for 30-90 minutes; and then heating the first mixture at 80-95° C. for 30-90 minutes to fractionize the first mixture to form an aqueous phase and an oil phase; (3) collecting an aqueous solution from the aqueous phase; and (4) isolating nucleic acids from the aqueous solution. The method improves the efficiency and convenience of isolating nucleic acids from FFPE tissue samples.

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: 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: 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: Compositions and methods for the efficient extraction, enrichment and isolation of nucleic acids from fresh, fixed or fixed and embedded cells, tissues, biological materials and cellular source material.

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 is directed to a genomic DNA extraction reagent and method for improved extraction of DNA from biological tissue. The extraction reagent of the invention is mixed with disrupted biological tissue to form a DNA extraction solute which is incubated in a DNA extraction step. The extraction reagent includes an alkali component to maintain the DNA extraction solute at a pH of about 10 to 14 substantially throughout the extraction step. The extraction solute is centrifuged to clarify the supernatant. The supernatant containing the extracted DNA is diluted with a neutralizing buffer resulting in a high throughout method of generating high quantities of high quality DNA. Major PCR inhibitors are managed with the unique chemical combinations of the DNA extraction reagent designed and optimized for extraction of DNA from plant tissue and cells.

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: Methods for extracting high quality nucleic acids from a heterogenous collection of nucleic acid-containing materials from a biological sample are disclosed. The heterogenous collection of nucleic-acid containing materials may contain cells or microvesicles, or both. The extractions obtained by the methods described herein are characterized by high yield and high integrity, making the extracted nucleic acids useful for various applications in which high quality nucleic acid extractions are preferred, e.g., a diagnosis, prognosis, or therapy evaluation for a medical condition.

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: 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: 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: Methods for processing polynucleotide-containing biological samples, and materials for capturing polynucleotide molecules such as RNA and/or DNA from such samples. 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.

Abstract: The invention relates to a method for purification of nucleic acids, to a kit for performing the method according to the invention and to a new application of magnetic particles for purification of a biological sample. The method according to the invention comprises the following steps: a) accommodating of the sample in a first sample vessel in an aqueous solution and lysing of the sample under non-chaotropic conditions; suspending of first magnetic particles in the solution and inserting of the first sample vessel in a sample vessel holder, wherein the sample vessel is inserted in the annular interior space of a ring magnet associated with the sample vessel holder; separating of the solution from the magnetic particles; and isolating of the nucleic acids from the solution.

Abstract: The present invention provides a method of isolating nucleic acid from a sample, said method comprising contacting said sample with a detergent and a solid support, whereby soluble nucleic acid in said sample is bound to the support, and separating said support with bound nucleic acid from the sample. Where the method of the invention is used to isolate DNA, it may conveniently be couple with a further step to isolate RNA from the same sample.

Abstract: The present invention provides a method for the isolation of biological molecules by the adsorption of the molecules onto a mineral substrate in the presence of an appropriate mixture of salts and sufolane. Preferably, the biological molecules are nucleic acids. Compositions and kits for performing the process according to the invention are also provided.

Abstract: A method for purifying a protected oligonucleotide comprising the steps of: a) providing a solution of the protected oligonucleotide in at least one solvent A having a boiling point below the boiling point of a solvent B, heating the solution at a temperature of at least 30° C. and below the boiling point of the at least solvent A, adding solvent B until precipitation of a material is visible in the solution, said solvent B being an alcohol having 1 to 6 C-atoms or a diol having 2 to 6 C-atoms, allowing the solution to cool down under stirring until formation of a supernatant and a residue, removing the supernatant or b) providing solvent B, said solvent B being an alcohol having 1 to 6 C-atoms or a diol having 2 to 6 C-atoms, heating solvent B at a temperature above 30° C.

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 invention provides systems, methods and kits for the separation and/or purification of double-stranded and single-stranded nucleic acids from the same sample. The method includes first mixing a sample containing both double-stranded nucleic acid and single-stranded nucleic acid with a solution including a chaotropic salt and a non-ionic detergent to generate a mixture; then applying the mixture to a first mineral support for double-stranded nucleic acid to bind; and collecting the flow-through which contains unbound single-stranded nucleic acid. The method further includes diluting the non-ionic detergent of the flow-through, and applying the diluted flow-through to a second mineral support for the single-stranded nucleic acid to bind. Alternatively the flow-through can be mixed with a lower aliphatic alcohol prior to loading of the second column. The double-stranded and the single-stranded nucleic acids can be eluted from the mineral supports respectively.

Abstract: A method of isolating nucleic acids from a biological material, comprises applying the biological material on a substrate comprising one or more cell lysis reagents impregnated therein; applying a fluid to the biological material applied on the substrate; extracting the nucleic acids from the biological material applied on the substrate; and collecting the extracted nucleic acids in a substantially intact form, wherein the collected nucleic acid has a molecular weight greater than or equal to 20 kb.

Abstract: A device, a system, and a method for isolating biomolecules from biological materials are provided. The device comprises a quartz-based solid phase extraction matrix comprising one or more reagents impregnated therein; and an electroosmotic pump (EOP) operationally coupled to the quartz-based solid phase extraction matrix to elute the nucleic acids, wherein the EOP comprises a plurality of electroosmotic membranes comprising one or more positive electroosmotic membranes and one or more negative electroosmotic membranes disposed alternatively and a plurality of electrodes comprising one or more cathodes and one or more anodes, wherein at least one cathode is disposed on one side of one of the membranes and at least one anode is disposed on another side of that membrane and at least one cathode or anode is disposed between a positive electroosmotic membrane and a negative electroosmotic membrane.

Abstract: The present invention relates to a method for the isolation and purification of nucleic acids by elution of nucleic acids from nucleic acid-containing samples, and biological materials, using a wash buffer comprising an alcohol having 1 to 3 carbon atoms and at least one further solvent selected from the group consisting of alkane diols and alkane triols having 2 to 6 carbon atoms, monocarboxylic acid esters and dicarboxylic acid diesters having 2 to 6 carbon atoms in the acidic component and 1 to 4 carbon atoms in the alcoholic component; (poly)ethylene glycols and ether derivatives and ester derivatives thereof, and poly(4-styrene sulfonic acid-co-maleic acid) sodium salt solution. The present invention further relates to a kit for carrying out the method of the invention.

Abstract: A solid matrix for the extraction, stabilization, and storage of nucleic acids is provided. At least one protein denaturant, and at least one acid or acid-titrated buffer reagent are impregnated in a dry state therein the matrix; and the matrix is configured to provide an acidic pH on hydration. The matrix is configured to extract nucleic acids from a sample and stabilize the extracted nucleic acids, particularly RNA, in a dry format under ambient conditions for a prolonged period of time. Methods for collecting and recovering the nucleic acids stored in the dry solid matrix are also described.

Abstract: The current disclosure provides methods and kits for isolating nucleic acid from an environmental sample. The current methods and compositions further provide methods for isolating nucleic acids by reducing adsorption of nucleic acids by charged ions and particles within an environmental sample. The methods of the current disclosure provide methods for isolating nucleic acids by releasing adsorbed nucleic acids from charged particles during the nucleic acid isolation process. The current disclosure facilitates the isolation of nucleic acids of sufficient quality and quantity to enable one of ordinary skill in the art to utilize or analyze the isolated nucleic acids for a wide variety of applications including, sequencing or species population analysis.

Abstract: The present disclosure provides methods for isolating substantially pure and undegraded DNA from biological material via a solid support. The methods may use a DNA lysing solution that comprises a surfactant, a DNA-complexing salt (e.g., a lithium salt), and a buffer. Alternatively, the methods may use a solid support pre-treated with the DNA lysing solution.

Abstract: A method of extracting a nucleic acid from a microvesicle, the method comprising treating the microvesicle with a composition comprising a detergent and an aprotic solvent to extract a nucleic acid from the microvesicle.

Abstract: DNA is extracted from epithelial cells and other cells without cell walls, by use of a DNA extraction buffer that contains a C1-C4 alcohol, a cell lysis detergent, and a buffering agent maintained at a slightly basic pH. The cells are immersed in the buffer and gently agitated at ambient temperature, and DNA extraction and precipitation occur in thirty minutes or less, and often in five minutes or less.

Abstract: The present disclosure generally relates to solid matrices for the extraction, stabilization, and storage of nucleic acids, particularly RNA, in a dry format under ambient conditions for a prolonged period of time. Methods for extracting, collecting, and recovering nucleic acids from the solid compositions are also described.

Abstract: The present invention provides a method of isolating nucleic acid from a sample, said method comprising contacting said sample with a solid support, preferably magnetic beads, in the presence of an ethylene glycol multimer consisting of from 2 to 70 ethylene oxide monomers, preferably tetraethylene glycol, whereby soluble nucleic acid in said sample is bound to the surface of the support, and separating said support with bound nucleic acid from the sample. Kits for performance of the invention are also provided.

Abstract: The present invention concerns the use of methods and compositions for the isolation of small RNA molecules (100 nucleotides or fewer), such as microRNA and siRNA molecules. Such molecules are routinely lost in commonly used isolation procedures and therefore the present invention allows for a much higher level of enrichment or isolation of these small RNA molecules.

Abstract: The invention includes a method for separating polynucleotides, such as DNA, RNA and PNA, from a solution containing polynucleotides by reversibly and non-specifically binding the polynucleotides to a solid surface having a functional amide group-coated surface. The materials containing a solid surface can be in the form of microparticles, fibers, beads, membranes, test tubes, pipette tips or microwells and can further comprise a magnetic core portion. The pH, salt and concentration of crowding reagent, such as polyethylene glycol or alcohol, of the solution is adjusted to levels which result in nucleic acid binding to the solid surface. The magnetic microparticles with bound polynucleotides are separated from the solution under mild alkaline conditions and the nucleic acids are eluted from the magnetic microparticles. Solutions having different nucleic acid concentrations can be normalized by restricting the availability of the solid phase surface.

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: There is provided a method of extracting a nucleic acid analyte from a cell or virus in a sample chamber, comprising a) adding disruption beads comprising external silica or glass to the sample chamber; b) agitating the disruption beads within the sample chamber to disrupt the cell; c) adding binding particles comprising external silica or glass to the sample chamber in the presence of a chaotropic agent; d) contacting the contents of the sample chamber with a removal device with which the binding particles reversibly associate; and e) separating the removal device and associated binding particles from the sample chamber, thereby removing the nucleic acid analyte from the sample. There are also provided apparatus and kits for use with the method.

Abstract: A formulation containing guanidine thiocyanate or guanidine hydrochloride together with acetamide, one or more acetamide derivatives, or a combination of acetamide and one or more acetamide derivatives is provided with methods to use the formulation to purify one or more nucleic acids contained in a medium.

Abstract: Disclosed are a chaotropic agent; a reagent including a chaotropic agent and a lithium salt; a reagent kit including a chaotropic agent; a chaotropic agent, a reagent, a reagent kit, and a method for isolating a nucleic acid by use of a magnetic cellulose material; a method for binding a nucleic acid to a magnetic cellulose material; a method for isolating a nucleic acid; and a method for purifying a chromosome DNA. It is required that each of the chaotropic agents, the reagents, and the reagent kits works with at least one solid-phase, magnetic cellulose-containing carrier to isolate a nucleic acid from non-nucleic acid substances. In addition, each chaotropic agent includes an alcohol substance and a substrate solution for adjusting the alcohol substance to an appropriate concentration and thereby promoting binding of the nucleic acid in a sample to the magnetic cellulose.

Abstract: A microfluidic apparatus, method, and associated applications utilize and apply to a formalin-fixed paraffin-embedded (FFPE) tissue sample and performing a liquid-liquid extraction to remove the paraffin from the tissue sample prior to a nucleic acid purification step. A microfluidic device includes a dedicated liquid-liquid extraction process vessel, a nucleic acid purification process component, and a nucleic acid amplification reactor. A liquid-liquid extraction and nucleic acid purification kit includes a microfluidic device capable of performing both a liquid-liquid extraction process and a nucleic acid purification process, including a dedicated liquid-liquid extraction process vessel, an immiscible liquid or a precursor phase thereof disposed in the vessel, a nucleic acid purification process component, a nucleic acid amplification reactor fluidically, and a supply of reagents suitable to enable the liquid-liquid extraction process and the nucleic acid purification process.

Abstract: The present invention is a method of separating nucleic acids using a solid phase capable of binding nucleic acids, such as magnetic glass particles, where binding of nucleic acids to the solid phase is enhanced by the presence of an ethylene-amine compound. The invention further includes a reaction mixture for isolating nucleic acids containing an ethylene-amine compound and kits for carrying out the method.