Abstract: This invention relates to methods for rapid nucleic acid purification from sources heavily contaminated with high particulate material, such as cellular debris, and solids, including suspended solids. In particular, this invention provides methods for rapid, quantifiable recovery and purification of nucleic acids from a variety of sources heavily contaminated with solids, such as small organisms, tissue samples, samples of blood found on soil, or samples of washing from foods, which are frequently difficult sources for nucleic acid isolation due to their propensity to clog filters and columns. A device and kit are also provided.

Abstract: The invention provides a novel method for the screening and for the identification of nucleic acid binding factors (NABFs) or nucleic acid binding elements (NABEs) that are differentially active between two phenotypically different cell lines (normal and modified); in a particular cell type before and after a given treatment; or between two completely heterologous cell lines. The invention also provides methods for the simultaneous analysis of the effect of given compounds on multiple DNA-protein interactions. It further provides for the analysis of compounds that are nucleic acid binding factor analogs and compounds that selectively bind cis-acting nucleic acids.

Abstract: The invention relates to a method for producing an aqueous extract rich in nucleic acids from a plant material, in particular from plant embryos or from seeds rich in DNA or in RNA, which consists in:

Abstract: The present invention relates to a method to isolate fully methylated unamplified eukaryotic DNA made of hundreds-of-thousand to billions of base pairs from a lysate. The eukaryotic tissue samples are digested with protolytic enzymes in buffer releasing cellular contents. Magnetically responsive functionalized solid particles are added to the lysate. Polyethylene Glycol and a high salt concentration are added to the mixture to disrupt the hydrogen bonding in the lysate. The eukaryotic genomic DNA binds to the functionalized microparticles and is washed a number of time with an alcohol mixture to remove and denature proteins. The genomic DNA is the eluted off the magnetically responsive functionalized solid particles and used in downstream reactions.

Abstract: A container assembly for storing and stabilizing a biological sample includes a container, a closure cap and a sample holder coupled to the closure cap and removably received in the container. The sample holder can be a basket-like device coupled to an inner face of the cap and includes a central cavity for receiving the sample and immersing the sample in the reagent in the container. The closure cap includes a body member with a dimension to displace a volume of air and reduce the head space to ensure that the sample holder is completely immersed in the reagent. The sample holder has a closure member for closing the open top end of the cavity.

Abstract: The present invention relates to an automatic DNA isolation apparatus which comprises liquid containers; numerous liquid flow passages; control valves; numerous syringes; numerous injection needles; transportation means; vacuum block; plate rack (31); numerous multi-well plate blocks; and process control unit which regulates the operation of said control valves for selection and transfer of said various liquids, the operation of said syringe for quantitative intake and discharge of said liquids, the transfer of said injection needle connected with said syringes, the sequential transportation of said multi-well plates arranged in said multi-well plate rack, the sequential operation of shaking block and said vacuum block.

Abstract: A nucleic acid derived from Bacillus thuringiensis contains a nucleotide sequence that encodes a polypeptide demonstrated to be toxic to fire ants.

Abstract: The invention relates to a method for removing endotoxins from nucleic acids. The endotoxins are removed by pre-incubating the nucleic acids in a salt-free detergent solution and subsequent anion exchange chromatography on a tentacle anion exchanger.

Abstract: A process for identifying genes coding for cellular constituents involved in regulating angiogenesis including culturing endothelial cells on an extracellular matrix protein according to at least four different types of conditions: a reference condition, an angiogenesis promoting condition, an angiogenesis inhibiting condition, and a control condition; isolating messenger RNAs stemming from cells cultured according to the different conditions; and comparing at the qualitative and/or quantitative level, different messenger RNA populations to identify messenger RNAs stemming exclusively or in a particularly elevated quantity from cell cultures under conditions stimulating and/or inhibiting angiogenesis, the messenger RNAs corresponding to the genes coding for the cellular constituents involved in regulating angiogenesis.

Abstract: A method for extracting nucleic acids from a biological material such as blood comprises contacting the mixture with a material at a pH such that the material is positively charged and will bind negatively charged nucleic acids and then eluting the nucleic acids at a pH when the said materials possess a neutral or negative charge to release the nucleic acids The nucleic acids can be removed under mildly alkaline conditions to the maintain integrity of the nucleic acids and to allow retrieval of the nucleic acids in reagents that are immediately compatible with either storage or analytical testing.

Abstract: A method for the indirect extraction of DNA greater than 300 kb in size from non-cultivatable organisms contained in an environmental sample is disclosed. The method comprises isolating the organisms from the sample and embedding the isolated organisms in a block of agarose where the organisms are subsequently lysed and the DNA subjected to a first alternating field electrophoresis to extract DNA fragments which are large in size and separate them from the cell debris. The first electrophoretic migration is followed by an enzymatic restriction step and additional electrophoretic migrations. The invention also encompasses DNA obtained by the method.

Abstract: The invention provides novel reagents, reagent kits, and methods for automated hybridization. More particularly, the invention provides reagents, reagent kits, and methods for automated in situ hybridization and automated hybridization on a microarray. The use of automated instruments for in situ hybridization and microarray hybridization dramatically reduces the amount of labor and time involved and also facilitates standardization of protocols and consistency between results.

Abstract: A method for simultaneous release and detection of nucleic acids from complex biological samples is described. The invention relates to the combined use of lysis buffers containing strong chaotropic agents such as guanidine thiocyanate to facilitate cell lysis and release of cellular nucleic acids and to the use of a novel type of bicyclic nucleotide analogues, locked nucleic acid (LNA) to detect specific nucleic acids released during lysis by nucleic acid hybridisation. In particular methods are described for the covalent attachment of the catching LNA-oligo. Novel methods for sample preparation of e.g. polyadenylated mRNA species are also presented. The invention further addresses reagents for performing the methods as well as reagents and applications of the method.

Abstract: A process and apparatus for isolating and purifying nucleic acids and other target molecules directly from blood, plasma, urine, cell cultures and the like by totally automated means, without centrifugation, aspiration or vacuum. After mixing a sample containing target molecules with a test reagent in an environmentally isolated compartment, target molecules are adsorbed onto a binding material and eluted in a small volume using an elution reagent. A preferred embodiment purifies nucleic acids and automatically detects target sequences from a sample of fresh blood.

Abstract: The present invention relates to a composition and to a method for extracting DNA. More specifically, the present invention relates to a composition and to a method to extract DNA from dried biological samples on solid substrates, including but not limited to, buccal smears, semen and especially blood. The method can be conducted in a single-tube. The DNA extracted in accordance with the present invention can be used for DNA amplification reactions, DNA sequencing, DNA restriction analysis and DNA hyridization.

Abstract: The present invention refers to a device and a procedure for the collection and initial preparation of tissue/blood or other sample of nucleated or DNA-containing cells or cell components for molecular genetic investigation. The invented device for the collection and initial preparation of samples of DNA-containing cells includes a sample receiving container and means for the collection of the sample, which is introduced into the sample receiving container after collection of the sample and seals this tightly. The sample receiving container has a base and side walls, is closed with a easily penetrable lid and has—in an area of the side walls of the container removed from the base—means to secure the introduced sample collection tool; in the container are substances to protect from DNA-degrading enzymes.

Abstract: A mechanical cell lysis technique involving the use of compaction protection technology to shield nucleic acids during mechanical lysis. Mechanical lysis is an efficient and widely used method of liberating the contents of microbial cells, but the shear sensitivity of large nucleic acids impairs the application of this technique to DNA purification. The invention uses compaction agents, small polycations that condense nucleic acids, to protect DNA from shear damage and allow mechanical lysis to be used in chromosomal and plasmid DNA purification. In addition to protecting DNA during lysis, compaction allows DNA to be pelleted with the insoluble cell debris, washed, and resolubilized to yield an enriched DNA product. Highly shear-sensitive nucleic acid molecules such as large plasmids and BACs can also be protected during lysis. An added benefit is that lysate viscosity is greatly reduced, allowing for reduced volumes compared to alkaline lysis.

Abstract: Particle aggregation of lipid:nucleic acid complex particles is prevented by incorporating a non-cationic lipid into lipid:nucleic acid complex particles containing a cationic lipid and a nucleic acid polymer. The non-cationic lipid is a polyethylene glycol-based polymer.

Abstract: The invention relates to compositions and methods for isolating nucleic acids from biological samples, including whole tissue. The invention also provides kits for isolating nucleic acids from biological samples.

Abstract: The invention describes a method for the isolation of components from samples, particularly large molecular weight DNA from biological samples. The method involves the application of controlled oscillatory mechanical energy to the sample for short periods of time of about 5 to 60 seconds to lyse the sample and release the component(s) from the sample, followed by standard isolation methods. In preferred embodiments, the method includes the use of a spherical particle for applying the mechanical energy.

Abstract: A method for isolating nucleic acid which comprises:

Abstract: Disclosed are methods and compositions for extracting nucleic acids from a biological sample. In particular, disclosed is a nucleic acid extraction solution together with methods using such a solution for extracting nucleic acid sequences from biological samples containing cells, cellular debris or both. The nucleic acid extraction solution contains a molecule having the formula R1O—CH2—CH2—OR2, wherein R1 and R2 independently are selected from the group consisting of hydrogen and an alkyl group.

Abstract: Disclosed is a method for removing polynucleotide from the skin. This polynucleotide can be used to detect dermatitis and distinguish an irritant reaction from an allergic reaction by characterizing the polynucleotide according to the polypeptide which it encodes. Additionally, provided are methods for non-invasive isolation of samples from the skin as well as kits for use in the methods provided herein.

Abstract: Intracellular material is released from bacterial, yeast, plant, animal, insect or human cells by the application of a low voltage such as 1 to 10 V to a suspension containing the cells. The conditions may be selected such that DNA released from the cells is electrochemically denatured so as to be available for use in an amplification procedure.

Abstract: The present invention concerns methods for purifying nucleic acids from whole cells in a sample, such as whole blood. Also provided is the use of a filter device in same.

Abstract: Methods are provided for the isolation of newly synthesized mRNA. Such methods involve the incorporation of biotinylated rNTP analogues into cellular mRNA, and separating biotinylated mRNA from unlabeled RNA. The methods provided herein may be used, for example, for gene discovery, drug screens and studies of the regulation of gene expression.

Abstract: The present invention relates generally to compositions, methods and kits for use in clarification and viscosity reduction of biological samples. More specifically, the invention relates to such compositions, methods and kits that are useful in the isolation of biological macromolecules from cells (e.g., bacterial cells, animal cells, fungal cells, viruses, yeast cells or plant cells) via lysis and one or more additional isolation procedures, such as one or more filtration procedures. In particular, the invention relates to compositions, methods and kits wherein biological macromolecules are isolated using a filter, where the pore size increases in the direction of sample flow. The compositions, methods and kits of the invention are suitable for isolating a variety of forms of biological macromolecules from cells. The compositions, methods and kits of the invention are particularly well-suited for rapid isolation of nucleic acid molecules from bacterial cells.

Abstract: The invention provides efficient methods for the isolation of centromeres from potentially any organism. The methods represents an advance over the prior art in that costly and labor intensive mapping programs are not required. Using the technique, methylated centromere DNA may be isolated from potentially any centromere in an organism. The technique is amenable to mass screenings employing use of arrays comprising libraries of DNA from a target species.

Abstract: This invention provides for a rapid and convenient method of simultaneous collection of both genomic and diagnostic information from a single sample on a bibulous pad by differential extraction of the diagnostic information from the genomic information. It is a surprising discovery of this invention that a PCR assay on the contents of the bibulous pad provides results comparable in reliability, specificity, and sensitivity to the best available serum (blood) based assays. The assays of this invention can be used to confirm each other, either by detecting the genomic information leading to the diagnostic information, or by detecting in the genomic information, a predisposition to a disease and confirming the presence of the disease through diagnostic testing.

Abstract: The invention includes reagents and methods for the isolation of nucleic acids. The reagents described herein contain a nucleic acid precipitating agent and a solid phase carrier. The reagents can optionally be formulated to cause the lysis of a cell. These reagents can be used to isolate a target nucleic acid molecule from a cell or a solution containing a mixture of different size nucleic acid molecules. The disclosed reagents and methods provides a simple, robust and readily automatable means of nucleic acid isolation and purification which produces high quality nucleic acid molecules suitable for: capillary electrophoresis, nucleotide sequencing, reverse transcription cloning the transfection, transduction or microinjection of mammalian cells, gene therapy protocols, the in vitro synthesis of RNA probes, cDNA library construction and PCR amplification.

Abstract: A method and system are provided for preserving nucleic acids in a bodily fluid, such as urine, blood, blood serum, and amniotic fluid. The preservative includes an amount of a divalent metal chelator selected from ethylenediaminetetraacetic acid (EDTA), [ethylenebis(oxyethylenenitrilo)]tetraacetic acid (EGTA) and 1,2-bis(2-aminophenoxy) ethane-N,N,N′,N′-tetraacetic acid (BAPTA), or salts thereof in the range of from about 0.001M to 0.1M; and an amount of at least one chelator enhancing component selected from lithium chloride, guanidine, sodium salicylate, sodium perchlorate, and sodium thiocyanate in the range of from about 0.1M to 2M.

Abstract: A method for obtaining molecules from a substantially intact single cell is disclosed. The method differs from prior methods that obtain a variable fraction of a cell's contents and therefore cannot quantitatively estimate the number of molecules in the cells. The present method comprises isolating and harvesting a substantially intact single cell from its organ tissue comprising the steps of subjecting a tissue mass to a dissociation method so that the cells are dissociated from the tissue to expose cell bodies or cell processes, contacting a dissociated cell with a device capable of collecting the cell from the tissue substantially intact, withdrawing device with the cell attached, and then isolating or detecting the molecules in the single cell.

Abstract: DNA containing nucleotide base mispairs can be isolated using a modified rolling circle amplification procedure. Specific Y-shaped adapters permit the selective circularization of these fragments with a complementary splint oligonucleotide. Rolling circle amplification is then carried out with a DNA polymerase. Rolling circle amplification can also be carried out using a mixture of DNA circles having different lengths. Genetic phase of linked DNA markers can be determined by selective amplification of one parental haplotype. DNA fragments can also be converted into a form that can be utilized as rolling circle amplification templates by ligation of hairpin forming adapters to the ends of the fragments. Two or more DNA polymerases can be used in a rolling circle amplification reaction. DNA polymerase III has special properties that improve rolling circle amplification.

Abstract: A method and kit for isolating nucleic acids from a nucleic acid containing starting material is disclosed, where the nucleic acids are released from the starting material and precipitated onto a trapping membrane. The method and kit may be used in the context of isolating genomic DNA from blood and isolating BACs from transformed bacterial cultures.

Abstract: Methods and compositions for isolation of genomic deoxyribonucleic acid from whole blood employ three distinct aqueous solutions, all of which are substantially free from chaotropic salts and organic solvents. Genomic DNA isolated according to the inventive subject matter is substantially free of polymerase inhibitors and of sufficient quality for quantitative PCR.

Abstract: Microorganisms are recovered from soil by suspending a soil sample containing the microorganisms in a buffer solution comprising an organic acid of 3 or more carbon atoms, and then separating the microorganisms into supernatant of the suspension. Furthermore, the citrate buffer is a citric acid and a salt of citric acid. Also microorganisms are separated from the soil particles and recovered from the suspension by centrifugation or electrophoresis.

Abstract: The present invention is directed to methods of isolation of related polynucleotides harboring nucleic acid difference within a polynucleotide sample. The method will be useful in detecting and identifying alternative splicing events and corresponding splicing isoforms and to detect genomic DNA differences between genomes. The method according to the present invention is based on the use of a single-stranded trap. The single-stranded trap preferably involves the use of single-strand binding protein.

Abstract: A method and system are provided for preserving nucleic acids in a bodily fluid, such as urine, blood, blood serum, and amniotic fluid. The preservative includes an amount of a divalent metal chelator selected from ethylenediaminetetraacetic acid (EDTA), [ethylenebis(oxyethylenenitrilo)]tetraacetic acid (EGTA) and 1,2-bis(2-aminophenoxy) ethane-N,N,N′,N′-tetraacetic acid (BAPTA), or salts thereof in the range of from about 0.001M to 0.1M; and an amount of at least one chelator enhancing component selected from lithium chloride, guanidine, sodium salicylate, sodium perchlorate, and sodium thiocyanate in the range of from about 0.1M to 2M.

Abstract: An apparatus for isolating cell material from a tissue system and/or a liquid includes (1) a vessel open at the top, into which the cell material to be isolated can be introduced in the tissue system and/or the liquid; and (2) a separating device having a stamp-shaped configuration and including a flat separating disk having a peripheral edge fitting with the inner walls of the vessel in a fluid-tight manner and presenting at least one passage opening which is covered by a filter membrane, and which device can be inserted from the top into the vessel. The isolating disk pressurises the cell material inclusive of the tissue system and/or the liquid and acts thereupon with shearing forces by rotation. The cells and/or cell systems pass through the pores of the filter membrane whereas the residual tissue material is retained.

Abstract: A process for the isolation and purification of nucleic acids and/or oligonucleotides for use in gene therapy wherein said nucleic acids and/or oligonucleotides are isolated or purified from an essentially biological source, characterized in that

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 DNA contained in a biological sample, including: lysing a DNA-containing biological sample and forming a DNA-bound carrier by placing a lysing solution, including the DNA-containing biological sample, a salt, and a cationic surfactant, and having a salt concentration higher than the DNA precipitation inhibition-initiating concentration, into contact with a DNA-binding carrier to bind DNA to the DNA-binding carrier to form the DNA-bound carrier; separating the DNA-bound carrier from other components; dissociating the bound DNA from the DNA-binding carrier; and recovering dissociated DNA. By the method, DNA purified with no preliminary treatment of a biological sample can be recovered at a high yield.

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 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 to monitor gene expression levels. The tissue sample can be a tumor or other pathological tissue.

Abstract: In one aspect the present invention provides methods for isolating nucleic acid molecules from a cell, the methods comprising (a) contacting a cell with a solution comprising a biopolymer-degrading enzyme, provided that the biopolymer-degrading enzyme is not a nuclease, and (b) contacting the cell with a solution comprising a hydrophobic surfactant to yield a cell suspension comprising cell, biopolymer-degrading enzyme and hydrophobic surfactant, wherein the hydrophobic surfactant has a critical micelle concentration less than 3.0 mM and the concentration of the hydrophobic surfactant in the cell suspension is at least 0.05% (v/v). In another aspect the present invention provides isolated DNA preparations comprising at least 80% supercoiled DNA. In another aspect the present invention provides isolated nucleic acid preparations having an A260/230 ratio of at least 2.0.

Abstract: A process for the recovery of plasmids or other DNA from cells using a first filtration step to remove the cellular debris and other large cellular components and then an ultrafiltration step to capture the plasmids or other DNA on the surface of the ultrafiltration membrane where they may be recovered. An apparatus is also taught for enacting the process and comprises an upper microfiltration or coarse filtration membrane and a lower ultrafiltration membrane. The driving force may be the same for both filters or different and may be done sequentially or simultaneously.

Abstract: A method of protein removal is provided, which utilizes a protein digesting enzyme and a detergent that is compatible with ultrafiltration. The method is particularly suited for isolating trace amounts of nucleic acid from a solution that has high protein concentration. The recovered nucleic acid is free of protein that may interfere with downstream application such as nucleic acid quantification or diagnostic use. A kit suitable for use in the protein removal method is also provided.

Abstract: DNA-containing hair root cells are liberated from hair samples by soaking the hair root ends of the hair shafts in an aqueous solution consisting essentially of dithiothreitol, a surfactant such as sodium dodecylsulfate, a chelate such as ethylenediamine tetraacetic acid, an inorganic salt (NaCl), tris(hydroxymethyl) aminomethane and sufficient hydrochloric acid to make the pH of the solution approximately 8.

Abstract: A scalable alkaline lysis process, including procedures and devices for the isolation of large quantities (grams and kilograms) of plasmid DNA from recombinant E. coli cells. Effective, controllable, and economical operation, and consistent low level of host chromosomal DNA in the final plasmid product. Involves a series of new unit operations and devices for cell resuspension, cell lysis, and neutralization.

Abstract: A process for the isolation and purification of nucleic acids and/or oligonucleotides for use in gene therapy wherein said nucleic acids and/or oligonucleotides are isolated or purified from an essentially biological source, characterized in that said essentially biological sources are lysed, the fractions obtained are optionally freed or depleted from the remainder of said biological sources by per se known mechanical methods, such as centrifugation, filtration; the fractions thus treated are subsequently treated with affinity chromatographic material or with inorganic chromatographic material for the removal of endotoxins; followed by isolation of said nucleic acids and/or oligonucleotides on an anion exchanger which is designed such that DNA begins to desorb from the anion exchanger only at an ionic strength corresponding to a sodium chloride solution of a concentration higher by at least 100 mM than one corresponding to the ionic strength at which RNA begins to desorb from the anion exchanger material.