Abstract: Nucleic acid material can be effectively separated from a fluid by first contacting the fluid with a positively charged polymer which binds the nucleic acid material. Thereafter, the polymer, having the nucleic acid material bonded thereto, is contacted with a releasing agent which comprises a solution of an alkaline material and a glycol. The solution has a pH of no more than 12 and operates to release the nucleic acid material from the polymer under relatively low temperature conditions, typically no more than 50° C., and in particular instances, no more than 40° C. The glycol material may comprise a monomeric glycol such as ethylene glycol, propylene glycol, or the like, or it may comprise a polymeric glycol such as polyethylene glycol. Also disclosed is a novel positively charged polymer which may be employed in the separation process. This polymer comprises an acidified polyamine, such as polyethyleneimine which has been reacted with a nonacidified polyethyleneimine in a coupling reaction.

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 polyamidoamine (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: Provided herein is technology relating to isolating nucleic acids. In particular, the technology relates to methods and kits for extracting multiple DNA targets from human stool samples.

Abstract: A method and kit which allow the use of a discrete amount of a binding matrix to first purify nucleic acids from a medium under a first set of binding conditions wherein the amount of nucleic acid bound to the binding matrix is essentially independent of the amount of surface area of the definable amount of the binding matrix, followed by a second purification step wherein the nucleic acids are bound to a discrete amount of binding matrix under a second set of binding conditions wherein the amount of nucleic acid bound to the binding matrix is essentially dependent on the amount of surface area of the definable amount of the binding matrix, thus providing a discrete quantity of nucleic acid.

Abstract: Formalin fixation causes cross-linkage between nucleic acids and proteins and covalently modifies RNA. As a result, the molecules are rigid and may comprise subsequent RNA extraction. The invention provides a method for recovering RNA from formalin fixed paraffin-embedded tissue, including a short additional step of incubation with proteinase K after the first digestion step that makes a significant enhancement of the quality and quantity of the extracted RNA and subsequently, an improvement in the detection of gene expression is achieved. The method of the invention has the advantage of minimizing the number of manipulations, eliminating the need for potentially toxic solvents, and increasing significantly the amount of RNA recovered, and therefore the sensibility, when compared with previous methods.

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 application is directed to a method for performing a bisulfite reaction to determine methylation positions in a nucleic acid via treatment of the solid phase-bound nucleic acid with bisulfite, desulfonation and elution of the nucleic acid from the solid phase.

Abstract: The present invention is a kit and method for isolating and quantitating miRNA and to the use of such methods in the diagnosis and prognosis of disease.

Abstract: Disclosed are water-soluble ionic liquids suitable for promoting adsorption of nucleic acids to a solid phase. The use thereof, particularly methods for the isolation of nucleic acids from an aqueous solution, as well as kits for performing those methods are disclosed.

Abstract: The invention provides a method for the separation and purification of two or three cellular components selected from genomic DNA, RNA and proteins from a single biological sample. The method comprises generating an aqueous solution containing the cellular components by lysing cells with a lysis solution; contacting the aqueous solution with an ion exchanger for genomic DNA and RNA to bind to the ion exchanger; collecting the flow-through which contains unbound proteins; eluting RNA from the ion exchanger; and eluting DNA from the ion exchanger. For the purification of any two of the cellular components, one of the components is not collected. The invention also provides reagent kits for carrying out the methods.

Abstract: A formulation containing guanidine thiocyanate together with acetamide, one or more acetamide derivatives, or a combination of acetamide and one or more acetamide derivatives is used to purify one or more nucleic acids contained in a medium. In particular, a medium containing at least one nucleic acid is combined with a binding matrix and the formulation in order to cause the at least one nucleic acid to separate from its in vivo cellular environment and to bind to the binding matrix. The binding matrix with at least one nucleic acid bound thereto then is separated from substantially the rest of the combined medium and formulation, after which the at least one nucleic acid is eluted from the binding matrix to obtain the at least one nucleic acid in a substantially purified form. If different nucleic acids are to be selectively purified from a single medium, multiple binding matrices, each compatible with a different nucleic acid, can be used.

Abstract: The present invention relates to the providing of a method for preparing a stool sample that enables a nucleic acid in a stool to be stably preserved without requiring a complex procedure, a solution for preparing a stool sample, a stool collection kit used in that method, and a method for recovering and analyzing a nucleic acid in a stool using a stool sample prepared using the preparation method of the present invention. A method for preparing a stool sample according to the present invention is a method for preparing a stool sample being used for analyzing a nucleic acid contained in the stool, and is characterized in that a collected stool is mixed with a solution having a protease inhibitor as an active ingredient.

Abstract: The present invention relates to the providing of a method for preparing a sample from a nucleic acid-containing sample such as biological samples, where inhibitory substance's action against a enzyme reaction using a nucleic acid as substrate are decreased, a solution for preparing a sample used for the method, a stool collection kit used in that method, and a method for recovering and analyzing a nucleic acid in a nucleic acid-containing sample using a sample prepared using the preparation method of the present invention. A method for preparing a sample according to the present invention is a method for preparing a sample being used for analyzing a nucleic acid, and is characterized in that a nucleic acid-containing sample is mixed with a solution having one or more members selected from the group consisting of a polycation and a chelating agent as an active ingredient.

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: A method of isolation of nucleic acids from a biological sample of cells comprising a combination of a solid phase cell nuclei isolation procedure with a solid phase nucleic acid isolation method.

Abstract: The invention relates to a method for preserving samples and disrupting cells which can be employed for the preparation of the extraction of nucleic acids from live cells, cell aggregates and tissue samples, and to a kit for carrying out the method. The method includes the introduction of live cells, cell aggregates or tissue samples whose nucleic acids are to be extracted into an excess of water-miscible and volatile organic liquid. The saturation of the sample with the organic liquid has an advantageous effect on the extraction of the nucleic acids, which is subsequently carried out in an aqueous medium. The mechanical cell disruption is facilitated by the virtually complete removal of the water from the cells. The biomembranes are dissolved in the organic liquid. tRNA and other RNA fractions with a low degree of polymerization, which are capable of permeating across the cell wall, can be extracted selectively without mechanical disruption.

Abstract: The present invention provides a method of purifying RNA, including contacting a solid support with an acidic solution having a RNA-containing sample and a kosmotropic salt having a concentration of less than 1M, thereby binding the RNA to the solid support. According to the present invention, RNA is purified efficiently due to high RNA yield and low contamination by DNA. The present invention is particularly effective in purifying RNAs of 200 nucleotides or less.

Abstract: A method for separating and purifying RNA including the steps of passing a sample solution containing a nucleic acid, a washing solution and a recovering solution through a nucleic acid-adsorbing porous membrane to adsorb nucleic, adsorbing, washing and recovering, in which the nucleic acid adsorbing porous membrane is a porous membrane capable of adsorbing a nucleic acid by interaction involving substantially no ionic bond, and the sample solution is obtained by a process, comprising the steps of (I) injecting a test sample containing at least one of blood and leukocyte, and further containing an anticoagulant to a container, (II) adding a hemolytic agent to the container to obtain a leukocyte pellet, (III) adding a nucleic acid-solubilizing reagent to the leukocyte pallet to obtain a mixture solution and (IV) adding a water-soluble organic solvent to the mixture solution to obtain the sample solution containing the nucleic acid.

Abstract: Embodiments of the present invention provide methods and kits for purifying nucleic acids. In particular, embodiments of the present invention provide methods and kits for purifying nucleic acids through the use of magnetic particles in binding buffers.

Abstract: The present invention describes isolation of plasmid DNA from bacteria. The addition of dyes to the alkaline lysis based purification buffers (P1, P2, and P3) allows for improved visual monitoring of the steps of preparing a bacterial lysate filtrate coupled to filtration or spin-column chromatography. The method comprises the suspending of the bacterial cells with buffer P1 (suspension is red/pink); lysing the bacteria with buffer P2 (suspension goes from red/purple color to translucent/purple); precipitating cellular debris with buffer P3 (solution becomes yellow with debris suspension); centrifuging or filtering to product a lysate filtrate; binding the lysate filtrate to a DNA binding matrix; washing; and isolating the plasmid via chromatography. The yield and quality of plasmid DNA is improved due to more consistent lysis. Errors in buffer addition are reduced by visualizing the color as buffers are added and also of changes in color of the preparation at each step.

Abstract: A phenol-free method of isolating DNA from biological material includes homogenizing a biological material with a homogenization buffer to form a homogenate. Proteins and non-DNA organic molecules are extracted from the homogenate by mixing a first extraction buffer and a second extraction buffer with the homogenate. The first extraction buffer includes chloroform and an alcohol and the second extraction buffer includes a non-ionic protein solubilizer and an alcohol. DNA is precipitated from the mixture of homogenate, first extraction buffer and second extraction buffer and the DNA is recovered by sedimentation.

Abstract: Methods for cell lysis and purification of biological materials, involving subjecting a sample to high pressure. Also featured is an apparatus for practicing the methods.

Abstract: Disclosed are compositions for collecting, storing, and transporting populations of nucleic acids from biological specimens, and clinical, forensic, or environmental samples. Also disclosed are methods for using these compositions as one-step formulations for killing pathogens, inactivating nucleases, and releasing polynucleotides from other cellular components within the sample, and stabilizing the nucleic acids prior to further processing or assay. In particular embodiments, the invention provides a single, one-step, sample collection/transport/storage formulation containing a known quantity of a non-genomic, nucleic acid carrier molecule that serves as an internal reference control to monitor the fidelity of the collection/transportation medium, and measure the integrity of nucleic acids subsequently isolated and purified from the processed sample.

Abstract: The invention relates to a modified spin column for the isolation and purification of plasmid DNA. A pre-filtration disc is included in a traditional spin column. During plasmid DNA isolation, the lysate can be loaded directly to the modified spin column, eliminates the need to first remove the flocculants containing cellular debris. This results in a much shortened process. Variation of the invention includes a depth filter in between the pre-filtration disc and the main separation matrix. Also provided are kits for isolation of plasmid DNA including the modified spin columns.

Abstract: The invention provides methods for removing a contaminant or inhibitor from a nucleic acid-comprising sample, wherein the contaminant or inhibitor inhibits the amplification or hybridization of the nucleic acid in the sample, or inhibits an enzymatic reaction utilizing the nucleic acid in the sample, the method comprising the steps of: (a) providing a reaction mixture comprising the sample, a chaotropic agent, ammonium acetate or an equivalent, and a detergent, (b) isolating the nucleic acid and remaining contaminants and inhibitors from the reaction mixture in a supernatant; and (c) contacting the nucleic acid supernantant with a flocculant resulting in the further removal of the contaminant or the inhibitor from the supernatant. The invention also provides kits that comprise the components necessary to carry out the method.

Abstract: Chromatographic loading solutions for use in the purification of trityl-on oligonucleotides. The solutions comprise an antichaotropic ion, a chaotropic ion, an alkaline salt, and a polar protic solvent, all at particular concentrations. The solution is useful in purifying oligodeoxyribonucleotides and oligoribonucleotides having either ACE or TBDMS protective caps. Also, methods and systems for purifying trityl-on oligonucleotides comprising the chromatographic loading solution, a reversed-phase sorbent, and the oligonucleotides to be purified.

Abstract: The speed of a nucleic acid amplification reaction, such as the Polymerase Chain Reaction (PCR), can be increased by setting the temperature of heat sources above and below the desired denaturation, annealing, and extension temperatures. The reaction mixture is only contacted with the heat sources long enough for the desired temperatures to be reached.

Abstract: A method for selectively separating and purifying RNA from a mixture solution of nucleic acid containing DNA and RNA, wherein the method comprising the steps of: (1-a) adsorbing nucleic acid; (1-b) washing; (1-c) subjecting to a DNase treatment; (1-d) washing; and (1-e) desorbing the RNA from a nucleic acid-adsorbing porous membrane by a recovering solution, wherein in the step (1-c), a total amount of a DNase solution is 130 ?l or less per 1 cm2 of the membrane. And a method for selectively separating and purifying RNA or DNA, which comprises the steps of: (2-a) adsorbing nucleic acid; (2-b) washing by a washing solution; and (2-c) desorbing the nucleic acid from a nucleic acid-adsorbing porous membrane, wherein the washing solution contains a water-soluble organic solvent having a concentration of 50% by weight or less, and does not contain a chaotropic salt.

Abstract: The present invention provides methods for purifying nucleic acid molecules, wherein each method includes the steps of: (a) synthesizing nucleic acid molecules in a reaction mixture; (b) contacting the nucleic acid molecules with a proteinase for a period of time sufficient to degrade protein in the reaction mixture; (c) applying the nucleic acid molecules treated in accordance with step (b) to a size-limiting filter so that at least some of the nucleic acid molecules are trapped on the filter; and (d) washing the filter with a phosphate buffer having a pH in the range of from about 5.7 to about 8.5.

Abstract: The present invention relates to a phase isolation process for biomacromolecule components, after pretreating the substance to be isolated: 1) adding sufficient amount of buffer (one of the compound selected from ammonium sulfate, cesium chloride, sodium sulfate, and ethylenediamine tetraacetic acid) and homogenizing gently to carry out agglutination; 2) adding phase isolation solution (one compound or a mixture of more compound selected from the group consisting of ethanol, isopropyl alcohol, isobutyl alcohol and acetone) and homogenizing gently, centrifuging to form an upper phase and a lower phase to carry out phase isolation; wherein lipid is in the upper phase, protein precipitates in the phase middle, DNA plasmid, viral nucleic acid, mitochondrial DNA are in the lower phase, and total RNA precipitates in the lower phase; genomic DNA precipitates in the lower phase or in the phase middle together with protein.

Abstract: Methods for deprotecting and derivatizing oligonucleotides that are non-covalently bound to a solid support are described. The methods include providing a plurality of oligonucleotides linked to one or more hydrophobic separation functions, wherein the plurality includes protected oligonucleotides, precipitating the plurality of oligonucleotides on a hydrophobic solid support using an organic solvent to produce non-covalently immobilized oligonucleotides, and deprotecting the immobilized oligonucleotides.

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: Reagents, methods and kits for the purification of RNA from biological materials are provided.

Abstract: A method for isolating plasmids DNA from a DNA containing material which comprises plasmid DNA and genomic DNA, comprising extracting the plasmid DNA into a water-immiscible organic solvent, a chaotrope and water under conditions to denature the genomic DNA and recovering the plasmid DNA from the organic phase.

Abstract: A nucleic acid-bondable magnetic carrier of the present invention is a magnetic silica particle comprising a superparamagnetic metal oxide, wherein the magnetic silica particle has a specific surface of about 100 to about 800 m2/g.

Abstract: Method for isolating DNA contained in a biological sample. The method includes combining in a solution a DNA-containing biological sample, a salt, a cationic surfactant, and a DNA-binding carrier, the solution having a salt concentration higher than the DNA precipitation inhibition-initiating concentration, to lyse the DNA-containing biological sample and to bind DNA to the DNA-binding carrier while in the solution to form a bound DNA-carrier. The method also includes separating the DNA-bound carrier from other components. The method further includes dissociating the bound DNA from the DNA-binding carrier. The method still further includes recovering dissociated DNA.

Abstract: A method for isolating nucleic acid which comprises: (a) applying a sample comprising cells containing nucleic acid to a filter, whereby the cells are retained as a retentate and contaminants are removed; (b) lysing the retentate from step (a) while the retentate is retained by the filter to form a cell lysate containing the nucleic acid; (c) filtering the cell lysate with the filter to retain the nucleic acid and remove remaining cell lysate; (d) optionally washing the nucleic acid retained by the filter; and (e) eluting the nucleic acid, wherein the filter composition and dimensions are selected so that the filter is capable of retaining the cells and the nucleic acid. Additionally, there is provided a substrate for lysing cells and purifying nucleic acid having a matrix and a coating and an integrity maintainer for maintaining the purified nucleic acid.

Abstract: Provided herein is a method for separating nucleic acid from a test sample comprising the steps of contacting a test sample with a metal oxide support material and a binding buffer to form nucleic acid/metal oxide support material complexes, separating the complexes from the test sample; and eluting the nucleic acid from the metal oxide support material.

Abstract: The invention relates to a method for linking nucleic acid and/or glycosaminoglycan or glycosaminoglycan mimetics to a polar/hydrophilic material, characterized by contacting a nucleic acid and/or glycosaminoglycan and a polar/hydrophilic material with each other in the presence of a solution being 20 to 100 percent saturated with a non-chaotropic salt and removing said solution from the nucleic acid and/or glycosaminoglycan—polar/hydrophilic material.

Abstract: The present invention is directed to compositions and methods for removal of nucleic acid probes from sample nucleic acids, particularly when the sample nucleic acids are attached to a solid support. The invention also concerns methods of stripping and reusing nucleic acid blots.

Abstract: The present invention provides RNA extraction reagents, methods and kits that are especially useful for extracting RNA. The reagents, methods and kits of the present invention are especially useful for extracting RNA, for example, cytoplasmic RNA, from difficult materials, from plants, especially, difficult plant tissues, such as those containing phenolics, tannins, polysaccharides (such as starch) and resins. Comparative high yields are obtainable according to the present invention when compared to conventional reagents and methods. The RNA preparations obtained in accordance with the present invention are also of high quality as demonstrated by superior A260/280 results and by gel electrophoresis.

Abstract: The present invention provides RNA purification and/or analysis methods that use ammonium sulfate to mitigate or neutralize inhibitory effects of certain molecules. Exemplary inhibitory molecules that interfere with RNA function or analysis are those that bind to or cleave RNA, or stabilize RNA secondary structures.

Abstract: The present invention is a general method for inactivating or inhibiting ribonucleases. Ribonucleases are treated with a reducing agent and heat. RNA samples contaminated with ribonuclease may be treated with this method to protect them from degradation. The RNA may then be used directly in a variety of enzymatic reactions and molecular biology techniques. This method may also be applied to a variety of molecular biology reagents which may be contaminated with ribonuclease to protect an RNA from being degraded when incubated with the reagent.

Abstract: The present invention relates to prognostic methods which are useful in medicine, particularly cancer chemotherapy. The object of the invention to provide a method for assessing GST-pi expression levels in fixed or fixed and paraffin embedded tissues and prognosticate the probable resistance or sensitivity of a patient's tumor to treatment with platinum-based therapies by examination of the amount of GST-pi mRNA in a patient's tumor cells and comparing it to a predetermined threshold expression level. More specifically, the invention provides to oligonucleotide primer pair GST-pi and methods comprising their use for detecting levels of GST-pi mRNA.

Abstract: The present invention relates to methods for lyophilizing eukaryotic cells and isolating intact nucleic acids from such cells, and related kits for using the same.

Abstract: The present invention provides methods for isolating a defined quantity of DNA target material from other substances in a medium. The method may be carried out using a known quantity of a silica-containing solid support, such as silica magnetic particles, having a definable capacity for reversibly binding DNA target material, and DNA target material in excess of the binding capacity of the particles. The methods of the present invention involve forming a complex of the silica magnetic particles and the DNA target material in a mixture of the medium and particles, and separating the complex from the mixture using external magnetic force. The DNA target material may then be eluted from the complex. The quantity of DNA target material eluted may be determined based on a calibration model. The methods of the present invention permit isolation of DNA target material which is within a known quantity range.

Abstract: The present invention provides a process of extracting nucleic acid in a simple manner at a high efficiency with no use of any hazardous reagent from biological materials having required specific procedure for nucleic acid extraction, particularly from biological materials such as bacteria of the genus Mycobacterium; and a process of simultaneously carrying out the extraction and purification of nucleic acid in a simple manner from such biological materials existing in a biological sample such as feces having involved much difficulty in the purification of the nucleic acid. In the present invention, vigorous agitation of biological materials with fine particles in solutions containing chelating reagents or agitation of biological materials with aqueous solutions containing chelating reagents and quaternary ammonium salts, together with organic solvents and fine particles is performed.

Abstract: Methods are disclosed for rapid, reliable and simple isolation of RNA from formalin-fixed paraffin-embedded tissue samples. RNA purified in this manner can be used for quantitative measurement gene expression levels. The tissue sample can be a tumor or other pathological tissue.

Abstract: Methods are disclosed for rapid, reliable and simple isolation of RNA, DNA and proteins from formalin-fixed paraffin-embedded tissue samples. RNA purified in this manner can be used to monitor gene expression levels. The tissue sample can be a tumor or other pathological tissue.

Abstract: Methods for extracting DNA from a biological sample that result in a higher yield of target DNA than conventional methods. More particularly, methods for extracting DNA include exposing the biological sample to inhibitors of DNA degradation.