Abstract: Described is a method of isolating cell components, such as nucleic acids, from natural sources by filtering a sample of the digested natural sources such as cells or cell fragments. The method is characterized in that the sample is passed through a filter, the pore size of which decreases in the direction of flow of the sample through the filter.

Abstract: A method of isolating a biological material by a) providing a biological material which is bound to a porous matrix (C11), and b) compressing the matrix under conditions where the biological material is released from the surface of the matrix into an elution liquid. brings about the advantage that the emission of aerosols into the environment is greatly reduced. It is also possible to obtain highly concentrated and minute amounts of solution. The method is also easy to automate.

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: The present invention relates to a method for diagnosing schizophrenia, said method being based on the detection in vitro of the presence of the allele Ep of the microsatellite HUNTH01 in the gene TH. The invention also relates to the primers used for implementing said method.

Abstract: Complementary DNA encoding a 52 kDa form of a protein present in the human Ro/SSA ribonucleoprotein complex has been cloned. A lambda gt11 cDNA library made from human thymocyte mRNA was screened with serum from a SLE patient and two immunoreactive clones were isolated. These clones reacted with other patient sera which had anti-52 kDa Ro/SSA antibodies and with affinity purified anti-52 kDa Ro/SSA antibodies. Moreover, affinity purified antibodies eluted from fusion proteins of the isolated clones reacted only with the 52 kDa protein of lymphocytes in the Western blot. Ro/SSA RNAs were also precipitated with these affinity purified antibodies, further confirming that the clones encode a 52 kDa Ro/SSA antigen. The sequence differs from the previously reported 60 kDa Ro/SSA gene.

Abstract: A process for the isolation and purification of nucleic acids and/or oligonucleotides for use in gene therapy wherein nucleic acids and/or oligonucleotides are isolated or purified from an essentially biological source, characterized in thatessentially biological sources are lysed, the fractions obtained are optionally freed or depleted from the remainder of 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 byisolation of 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.

Abstract: A rapid mutational analysis method for mapping protein epitopes is disclosed. This method has been used to identify the binding sites for 16 anti-CD2 and anti-CD4 monoclonal antibodies. The powerful, rapid, and simple method of the present invention allows isolation of a very large number of mutants, and is applicable to any intracellular or surface protein for which a cDNA and monoclonal antibodies are available. The present method is especially useful in ligand binding site studies for the design of new ligands and drugs.

Abstract: Solutions and methods are disclosed for the effective, simple isolation/extraction of DNA, RNA and proteins from a single biological material sample, such as cells, tissues and biological fluids. The preferred solutions include effective amounts of a chaotropic agent(s), buffer, reducing agent, and may or may not include an organic solvent. Genomic DNA and total RNA can be isolated utilizing the solutions and methods of the invention in as little as 20 minutes, and proteins in as little as 30 minutes.

Abstract: Sequencing methods and methods for synthesizing DNA probes using mutant bacteriophage T4 DNA polymerases which have increased ability to incorporate modified nucleotides for the synthesis of long or short chains of complementary, modified, e.g., fluorophore-labeled DNA. In general, the mutant T4 DNA polymerases retain 3'.fwdarw.5' exonuclease activity; hence, reduction or elimination of 3'.fwdarw.5' exonuclease activity is not a prerequisite for efficient synthesis of a complementary fluorophore-labeled or other modified DNA. In fact, retention of 3'.fwdarw.5' exonuclease activity increases accuracy of DNA replication, because these exonucleases proofread or edit the product of DNA replication.

Abstract: The present invention relates to devices and methods for the collection, storage, and purification of nucleic acids, such as DNA or RNA, from fluid samples for subsequent genetic characterization, primarily by conventional amplification methods. The present invention can be used to collect, store, or purify nucleic acids from a treated whole blood source that has naturally occurring nucleic acid amplification inhibitors present, as well as added blood stabilization components that also inhibit nucleic acid amplification. More importantly, these nucleic acids can be released after collection or storage in a manner that enables them to be amplified by conventional techniques such as polymerase chain reaction. In particular, an absorbent material that does not bind nucleic acids irreversibly is impregnated with a chaotropic salt. A biological source sample is contacted with the impregnated absorbent material.

Abstract: This invention is directed to a novel method for PCR amplification wherein PCR is carried out at a higher pH than the pH widely used in the art. Specifically, the buffer solution is adjusted to pH 9.0 to 11.0 at 25.degree. C. Using the present invention, DNA amplification can be successfully carried out following a simple pretreatment. In the present invention whole blood is mixed with a hypotonic solution so that a selective lysis of red blood cells takes place. The residual leukocytes are then collected. The leukocytes are mixed with a polymerization agent, primers and other necessary reagents and PCR is carried out. When the PCR solution is placed at a high temperature for DNA denaturation, the leukocytes are lysed so that the leukocyte DNA is released and can access the primers and the other necessary reactions for PCR in the solution.Cell membranes and proteins are present in the PCR reaction solution due to the lack of a protein extractive step during the pretreatment.

Abstract: A method for the separation of a target molecule from a mixture is described. The method employs oil bodies and their associated proteins as affinity matrices for the selective, non-covalent binding of desired target molecules. The oil body proteins may be genetically fused to a ligand having specificity for the desired target molecule. Native oil body proteins can also be used in conjunction with an oil body protein specific ligand such as an antibody or an oil body binding protein. The method allows the separation and recovery of the desired target molecules due to the difference in densities between oil bodies and aqueous solutions.

Abstract: The invention features a method of isolating nucleic acid in a substantially purified form, including the steps of: a) contacting a biological sample which contains aggregated nucleic acid with a matrix comprising a solid hydrophilic organic polymer without an effective positive charge under conditions which permit the nucleic acid to bind to the matrix; and b) recovering nucleic acid from the matrix.

Abstract: Use of isopropanol in aqueous solutions for chromatographic isolation of nucleic acids for enhancing the transfection efficiency of the isolated nucleic acids in prokaryotic and eukaryotic cells.

Abstract: The invention relates to a cartridge for preparing purified nucleic acids obtained from a sample of cells, the cartridge comprises a filter tube disposed inside a dialysis tube, the ends of the tubes are mounted in endpieces that are interconnected by rigid uprights, the endpieces are designed to be connected to a device for injection and delivery of substances such as a sample of cells, reaction substances, and rinsing substances.

Abstract: Bacteria which will suppress fungus-induced potato disease under storage conditions have been screened and selected from soil samples. A method for isolating these antagonists, their use in controlling potato disease, and specific isolates which are inhibitory to potato dry rot disease under post-harvest conditions constitute the essence of the invention. The subject biocontrol agents are considered to be economically-feasible alternatives to chemical agents currently in use for this purpose.

Abstract: The present disclosure describes a method for probing the integrity of a Y chromosome utilizing multiplex PCR reactions which amplify specific regions of the human Y chromosome which have been linked to normal fertility in human males. The method is capable of detecting deletion mutations within the Y chromosome which are predictive of human male infertility. A kit containing reagents needed to practice the method is also disclosed.

Abstract: A process is disclosed for reducing or removing endotoxins from compositions containing therapeutic active substances extracted from natural sources by genetic engineering and/or biotechnology. For that purpose, the compositions are treated with chromatographic materials. The natural sources are disintegrated, the thus obtained fractions are, if required, centrifuged, filtered or treated using affinity chromatography methods, the fractions are pre-incubated in an aqueous salt solution and detergents, are treated with anion exchange materials, then washed with another salt solution. The active substances are eluted from the anion exchanger then further purified in a manner known per se.

Abstract: Nucleic acid probes and primers are described for detecting fungi that cause disease in humans and animals, as well as spoilage of food and beverages. These probes can detect rRNA, rDNA or polymerase chain reaction products from a majority of fungi in clinical, environmental or food samples. Nucleic acid hybridization assay probes specific for Acremonium sp., Aspergillus clavatus, Aspergillus flavus, Aspergillus fumigatus, Aspergillus glaucus, Aspergillus nidulans, Aspergillus niger, Aspergillus ochraceus, Aspergillus terreus, Aspergillus unguis, Aspergillus ustus, Beauveria sp., Bipolaris sp., Blastoschizomyces sp., Blastomyces dermatitidis, Candida albicans, Candida glabrata, Candida guilliermondii, Candida kefyr, Candida krusei, Candida lusitaniae, Candida parapsilosis, Candida tropicalis, Chrysosporium sp., Cladosporium sp., Coccidioides immitis, Cryptococcus neoformans var gattii serotype B, Cryptococcus neoformans serotype A, Cryptococcus laurentii, Cryptococcus terreus, Curvularia sp., Fusarium sp.

Abstract: A method of producing a vaccine comprising providing a quantity of the disease-causing entity, treating the quantity to remove or disrupt a majority of the nucleic acid present in the quantity sufficient to render the quantity non-infective while maintaining the structure of the proteins and glycoproteins on the surface of the entity to render the vaccine protective. A method of producing a vaccine to human pathogenic retroviruses comprising the steps of partially solubilizing a quantity of the virus to produce a suspension, adding a proteinase to the suspension to release viral nucleic acids from the viral coat proteins, treating the suspension to disrupt or remove sufficient viral nucleic acids to render the suspension substantially or completely non-infective and verifying the sufficient disruption or removal of viral nucleic acids from the suspension.

Abstract: A plastic is recovered from microorganisms containing it by chemically solubilising non plastic material with an oxidising agent in the presence of a chelating agent.

Abstract: A method of recovering a biopolymer from solution involves the use of magnetically attractable beads which do not specifically bind the polymer. The beads are suspended in the solution. Then the polymer is precipitated out of solution and becomes non-specifically associated with the beads. When the beads are magnetically drawn down, the polymer is drawn down with them. The polymer can subsequently be resolubilized and separated from the beads.

Abstract: The invention relates to a cellular protein which is specific and has high affinity for nucleic acid sequences characteristic of an intact TAR RNA loop sequence of the HIV LTR TAR region. The invention also relates to a protein preparation having a protein of about 185 kD that is isolated from a mammalian cell nuclear extract preparation, most specifically a HeLa cell extract that is purified between 1,000-10,000 fold. The protein of about 185 kD is shown to regulate HIV viral gene expression by binding a TAR RNA region of an HIV LTR template, in the presence of a cofactor fraction (including at least a -100 kD cofactor), and a tat protein. A route for the development of immunodiagnostics for AIDS and related disorders may also be provided given the specific and high affinity of TRP-185 for HIV RNA.

Abstract: Microsatellite nucleotide repeat sequences present in DNA isolated from a biological sample are used to determine the origin of a biological sample by a process including the steps of isolating nucleic acid from a biological sample, determining the interspersion pattern of repeats of a nucleotide sequence selected from the group consisting of di-, tri-, and tetra- microsatellite nucleotide repeat sequences in the DNA, and comparing the determined interspersion pattern with known interspersion patterns of the nucleotide sequence in selected mammalian species.

Abstract: A method of recovering a first material, e.g. plasmid DNA, from a solution containing pre-precipitated second material, e.g. genomic DNA and cellular debris, by precipitating the first material in the presence of magnetically attractable beads which become non-specifically associated with the newly formed precipitate, using a magnet to draw down the beads and the newly formed precipitate, and removing the supernatant.

Abstract: The present invention provides an improved method for the preparation of ribonucleic acid (RNA) samples. This method utilizes heat and guanidinium thiocyanate treatment of samples followed by alcohol precipitation and centrifugation to prepare RNA samples with a high degree of sensitivity, reliability, and ease of use. Importantly, the prevent invention provides a method in which RNA samples may be prepared so as to conserve RNA preservation and precipitation reagents and time. The samples so prepared are readily amplifiable and may be used for other purposes as well.

Abstract: A device and a process for isolating nucleic acids by lysing intact cells and removing nucleic acids emerging from the lysed cells by the following steps:a) the cells are immobilized in a porous matrix, with the size of matrix voids being in the range of the type of cell to be lysed;b) the cells are lysed;c) the nucleic acids are fixated on the matrix surface, and subsequentlyd) are eluted.

Abstract: The invention relates to the identification of the molecular basis of supravalvular aortic stenosis (SVAS) and Williams syndrome. More specifically, the invention has identified that elastin causes or is involved in the pathogenesis of SVAS and Williams syndrome. Molecular variants of the elastin gene contribute to SVAS and Williams syndrome. The analysis of the elastin gene will provide an early diagnosis of subjects with SVAS and Williams syndrome. The diagnostic method comprises analyzing the DNA sequence of the elastin gene of an individual to be tested and comparing it with the DNA sequence of the native, non-variant elastin gene. In a second embodiment, the elastin gene of an individual to be tested is screened for mutations associated with SVAS or Williams syndrome. Presymptomatic diagnosis of SVAS and Williams syndrome will enable practitioners to prevent vascular obstruction using existing medical therapies like beta adrenergic blocking agents.

Abstract: A method is disclosed for diagnosing Hereditary Neuropathy with Liability to Pressure Palsies (HNPP). A submicroscopic deletion of about 1.5 million basepairs on chromosome 17p11.2 is associated with the disorder in three unrelated pedigrees. The deletion includes all the markers known to map within the Charcot-Marie-Tooth type 1A (CMT1A) duplication. The method involves detecting the presence or absence of the deletion in DNA extracted from a patient sample. The deletion may be detected by Southern analysis or fluorescence in situ hybridization analysis (FISH). Sequences or probes that may be used to detect the deletion are provided, as are components of a kit for diagnosing HNPP.

Abstract: Compositions and methods for isolating nucleic acids from biological tissues and cells and for tissue/cell solubilization for other molecular biological uses, wherein the compositions comprise, in part, novel combinations of chaotropic agents and aromatic alcohols which act synergistically to effect better tissue/protein solubilization. The inventive compositions further include aprotic solvents for deactivation of ribonucleases and denaturization of proteins, as well as detergents for enhancing cell lysis and nucleoprotein dissociation. The inventive methods also comprise the use of a centrifuge, a solid-support matrix, and a microporous membrane for final isolation of the precipitated nucleic acids, resulting in high yield and purity of the precipitated nucleic acid.

Abstract: The present invention relates to modified glass fiber membranes which exhibit sufficient hydrophilicity and sufficient electropositivity to bind DNA from a suspension containing DNA and permit elution of the DNA from the membrane. Generally, the hydrophilic and electropositive characteristics are expressed at the surface of the modified glass fiber membrane. Preferred modified glass fiber membranes of the present invention include glass fiber membranes that have been modified by treatment with trifluoroacetic acid (TFA), BCl.sub.3, SiCl.sub.4, NaOH, F.sup.-, AlCl.sub.3 alone or in combination, with or without water. The modified glass fiber membranes of the present invention are particularly useful in processes for purification of DNA from other cellular components.

Abstract: A method for observing and determining the size of individual particles and for determining the weight distribution of a sample containing particles of varying size, which involves placing a deformable or nondeformable particle in a medium, subjecting the particle to an external force, thereby causing conformational and/or positional changes, and then measuring these changes. Preferred ways to measure conformational and positional changes include: (1) determining the rate at which a deformable particle returns to a relaxed state after termination of the external force, (2) determining the rate at which a particle becomes oriented in a new direction when the direction of the perturbing force is changed, (3) determining the rate at which a particle rotates, (4) measuring the length of a particle, particularly when it is at least partially stretched, or (5) measuring at least one diameter of a spherical or ellipsoidal particle.

Abstract: Methods useful in purifying nucleic acids from whole cell samples are described. This is accomplished by adding one or both of a non-phenyl group containing non-ionic detergent or a cross-linked, polycarboxylic acid. Kits which can be used in the methods are also described.

Abstract: A method of recovering a biopolymer from solution involves the use of magnetically attractable beads which do not specifically bind the polymer. The beads are suspended in the solution. Then the polymer is precipitated out of solution and becomes non-specifically associated with the beads. When the beads are magnetically drawn down, the polymer is drawn down with them. The polymer can subsequently be resolubilized and separated from the beads.

Abstract: This invention relates to a process for the amplification of nucleic acids in the form of DNA or RNA from blood samples by means of an enzymatic amplification method, characterized in that no preparation of the blood sample otherwise necessary to prepurify the nucleic acid to be amplified is performed and the proportion of the sample in the reaction mixture for the amplification process is greater than 5 volume % if a specific amount of salt is present in the reaction mixture. Depending on the proportion of blood sample and its salt contribution of monovalent and/or bivalent ions, the salt concentration in the reaction mixture in which the amplification is performed is, where applicable, adapted to the enzyme requirements by the use of an appropriately concentrated salt solution.

Abstract: The invention provides a method for determining the genomic sex of various salmonids (family salmonidae). In particular, the invention provides the nucleic acid sequence of a pseudogene, designated GH-.PSI., which is linked to a sex determining locus on the Y chromosome, and may be used as a marker for determination of the sex of the fish.

Abstract: An apparatus and method for the preparation of large DNA molecules from cells which have been cast in agarose. Specifically, the invention provides for a processing chamber that allows agarose plugs to be molded and processed within the same apparatus. This greatly reduces the amount of manipulation required of such DNA samples and reduces the loss of material due to agarose plug breakage. The mold has a filling port for agarose and a slot for at least one retainer for preventing the molten agarose from exiting through openings in the mold through which DNA processing solutions later access the molded agarose plugs.

Abstract: A method of separating cell nuclei from cells comprises: treating a fluid containing whole cells so as to selectively lyse the cytoplasmic membrane together with a small proportion of the nuclear membranes but leaving a large proportion of the cell nuclei intact; applying the treated fluid to a membrane whereby a mesh of DNA from the lysed nuclei is formed on the surface and captures intact cell nuclei. A device for use in the method is described.

Abstract: The present invention relates to a device method and kit for the convenient and rapid isolation and purification of nucleic acids, proteins, peptides, carbohydrates and oligosaccharides from heterogeneous biological samples. The device comprises a membrane assembly comprised of layers of microporous, polymeric membranes functionalized with ion-exchange groups. The device is reusable for like samples, relatively inexpensive compared to currently available separation techniques and is disposable, thereby avoiding cross-contamination of biological samples.

Abstract: A novel rapid mutational analysis method for mapping protein epitopes is disclosed. This method has been used to identify the binding sites for 16 anti-CD2 and anti-CD4 monoclonal antibodies. The powerful, rapid, and simple method of the present invention allows isolation of a very large number of mutants, and is applicable to any intracellular or surface protein for which a cDNA and monoclonal antibodies are available. The present method is especially useful in ligand binding site studies for the design of new ligands and drugs.

Abstract: Alkaline proteases derived from specific bacteria of the species Bacillus proteolyticus have enhanced stability and improved washing ability when blended in general detergents. Also disclosed are new bacteria producing these alkaline proteases. Additionally, there is also disclosed a process for the production of the alkaline proteases which comprises cultivating new bacteria and detergent compositions containing these alkaline proteases.

Abstract: The invention involves the supercritical or near-critical fluid disruption of microbial cells and extraction of intracellular components. First, a solvent that is a gas at ambient conditions and that has a critical temperature of between 0.degree. and 100.degree. C. is selected. This solvent is brought to near-critical pressure or higher and to near-critical temperature. The solvent then is combined with a slurry of cells to saturate the cells with the solvent under the prescribed conditions. Next, the pressure is released to cause a pressure drop which results in partial disruption of the cell membrane and release of solvent and other materials from the cell. Novel apparatus and associated methods are provided for carrying out the foregoing process continuously.

Abstract: Methods and compositions for improved liquification of mucoid secretion specimens by treatment with a disulfide bond reducing agent and a DNA digestion agent, and for improved concentration of selected bacterial species from such specimens, are disclosed. The disclosure has applicability to bacterial assay techniques including nucleic acid hybridization, culture and stain techniques.

Abstract: A cartridge for preparing nucleic acids such as DNA from cells comprises two end rings (14) enabling it to be mounted in sealed manner in a tube (10), two dialysis membranes (16) delimiting a dialysis enclosure (18) between them, a filter (20) for retaining cell nuclei dividing said enclosure (18) into two separate compartments, and means (24) for feeding substances into one of the compartments, means (26) for extracting substances from the other compartments, and means (28, 30) for feeding a dialysis liquid into the tube outside the cartridge, or for causing it to flow therethrough. The invention also provides apparatus, and a method using said cartridge.

Abstract: A novel method for the isolation of high molecular weight DNA from plants, yeast bacteria, and animal cells or tissue employs xanthate forming compounds, such as sodium/potassium ethyl xanthogenate. The procedure does not require deproteination and yields clean DNA that is suitable for both PCR and Southern blotting. It can be utilized on a small scale without homogenizing the tissue. These features also facilitate automated screening of tissue samples, one of the labor-intensive techniques in molecular biology.

Abstract: This invention relates to an apparatus for automatic purification of extra-chromosomal DNA from a cell suspension in a container. The apparatus comprises a device for introducing the cell suspension from a container into a chamber; a device for agitating the contents of the chamber; a device for introducing a lysing solution to the chamber to lyse the cells therein; a device for introducing a precipitating solution into the chamber to precipitate chromosomal DNA and, optionally, proteins and cellular debris therein; a device for feeding the liquid contents of the chamber through a filter to a collecting system; a device for purifying extra-chromosomal DNA from the contents of the collecting device; a device for introducing a dissolving solution into the chamber to dissolve the precipitate remaining therein, and a device for feeding the dissolved precipitate to waste.

Abstract: An optical recording medium having a recording film which comprises a phthalocyanine type coloring matter and a light absorbing agent having an absorption in the wavelength region corresponding to the laser recording wavelength.Such an optical recording medium is excellent in light resistance, is capable of providing a clear pit having a neat configuration at the time of recording and is capable of providing a sufficient reproduction characteristic.

Abstract: Methods and compounds for the preparation of nucleic acid samples. Polymerase inhibitors are treated so as to allow for amplification of nucleic acid from biological sources, including whole blood.

Abstract: The present invention provides a method for rapidly obtaining substantially pure DNA from a biological sample containing cells. The method involves gently lysing the membranes of the cells to yield a lysate containing genomic DNA in a high molecular weight form. The lysate is moved through a porous filter to selectively trap the high molecular weight DNA on the filter. The DNA is released from the filter using an aqueous solution to form a solution containing substantially purified DNA, from which the DNA may analyzed or recovered.

Abstract: A biochemical procedure for identification and characterization of cells in a biopsy or sample of a body fluid. The method can be used to determine cell type, i.e. epidermal, neuronal; tissue of origin, i.e. breast tissue, liver tissue; and degree of abnormality. The procedure can also be used to make antibodies and hybridization probes to detect cell or tissue specific antigens and nuclear matrix associated nucleic acids in cellular material and body fluids.The procedure is based on the isolation and analysis of the components of a specific subcellular protein fraction referred to here as the "nuclear matrix". The nuclear matrix includes proteins and nuclear matrix associated DNA specific to different cell types. These proteins and nucleic acids are altered or new ones expressed as a result of viral infection, genetic defects or malignancy.