Abstract: A process for producing and purifying peptides and for producing peptide/protein antigens for antibody production is described. The process utilizes fusion proteins and specifically fusion proteins in which the fusion protein carrier segment includes an amino acid sequence at least about 65 amino acids long and in which the amino acid sequence does not contain negative or positive charged side chains of amino acids.

Abstract: Solutions containing nucleic acids are treated with an alkaline protease to digest proteins such as nucleases that degrade the nucleic acids. In the isolation of nucleic acids, a biological sample containing nucleic acids is suspended in a solution containing water, buffer and chelating agent, the pH of the solution is adjusted to at least about 10 by adding a solution of sodium hydroxide and anionic detergent, an alkaline protease is incubated in the solution until nucleases are degraded, the pH of the solution is lowered to reduce activity of the alkaline protease by adding a solution having a pH between 3.5 and 4.5 and the alkaline protease is heat inactivated. Lowering of the pH may produce a cloudy solution which is cleared by centrifuging. Nucleic acids are isolated from the cleared solution by alcohol precipitation, or by using paramagnetic particles or a resin matrix containing silica particles. A chaotropic salt can be used to reversibly bind DNA to the resin matrix.

Abstract: Site-directed in vitro mutagenesis is obtained utilizing a specially-engineered plasmid vector. The vector is engineered to contain an inactivated genetic marker which marker is capable of being reverted to functional expression. Additionally, the vector contains a second genetic marker, a polylinker region and an f1 replication origin.

Abstract: A first embodiment of the method is for analyzing the amount of methionine sulfoxide in a protein sample and includes the steps of contacting a protein solution with methionine sulfoxide reductase in the presence of a reducing reagent bearing a covalently-linked reporter tag, whereby the reducing reagent is oxidized. The oxidized reducing reagent formed, which is in proportion to the amount of methionine sulfoxide in the sample, is then quantified. A second embodiment of the method is for analyzing the amount of disulfide linkages in a polypeptide or protein sample. It proceeds in the same fashion as above, but in the absence of any enzyme. A novel fluorescently-labeled reducing agent, and kits to practice the method are also disclosed.

Abstract: Novel purified agarase enzymes from Flavobacterium sp. strain NR19 and cloned genes encoding the agarase enzymes are disclosed. Transformed host cells which express the novel agarase enzymes in isolatable quantities are also described. Also disclosed are antibodies specifically reactive with the novel agarases.

Abstract: A system is provided for filtering a substance from a solution for subsequent analysis of the substance. The system has a manifold and a removable cover capable of latching onto a base defining an interior of the manifold. Apertures are formed in the cover capable of receiving columns having a filter for filtering the substance from the solution as solution is drawn through the columns into the manifold. The columns may be integrally formed to provide a set of columns. As a result, an array of apertures can receive a number of sets of columns to conduct the filtering process.

Abstract: The present invention is directed to the simultaneous amplification of multiple distinct genetic loci using PCR or other amplification systems to determine in one reaction the alleles of each of the loci contained within the multiplex.

Abstract: The present disclosure discloses 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 for isolating nucleic acids from agarose, and particularly regular agarose, is described wherein the agarose is augmented with a chaotropic substance and hydrolyzed by a novel purified agarase enzyme from Flavobacterium sp. strain NR19.

Abstract: Methods and compositions are provided for improved kinetics of light production from luciferase activity in beetle luciferase-luciferin reactions utilizing a luciferase inhibitor. Thus, the invention provides methods and compositions for assaying samples for the presence of a beetle luciferase in conjunction with a luciferase inhibitor.

Abstract: The present invention provides compositions and methods for isolating nucleic acids with lengths greater than about 50 bases, from cells, gels, solutions and other media, in which nucleic acids occur in vivo or in vitro. The compositions of the invention are mixtures of the silica materials silica gel and glass particles, particularly glass microfibers; such mixtures combined with chaotropic salts, such as guanidinium chloride or guanidinium thiocyanate; and suspensions of such mixtures in aqueous solutions of chaotropic salts. In the methods of the invention, an aqueous solution comprising nucleic acid is mixed with an aqueous solution of chaotropic salts and the resulting solution is contacted with a mixture of the silica materials, whereupon the nucleic acid in the solution binds to the silica materials. The chaotropic salts and components, other than the nucleic acid adsorbed to the silica materials, from the aqueous solution treated by the method of the invention are washed from the silica materials.

Abstract: The presently disclosed invention is drawn to an automatic luminometer apparatus capable of measuring two distinct luminescent reactions from within a single, non-compartmentalized sample container. The present apparatus may be dimensioned, configured, and programmed to automatically perform dual-reporter luminescent assays using multi-well sample plates, such as 96-well microtiter plates.

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: The present invention is directed to an assay system, a kit and a process for detecting at least one short tandem repeat sequence from DNA at a specific locus utilizing an allelic ladder containing at least two short tandem repeat sequences of the same lengths as two or more known alleles for the locus.

Abstract: Methods, kits, and reagents for conducting site-specific mutagenesis of single or double-stranded nucleic acids which utilizes novel antibiotic resistance conferred by a mutated antibiotic resistance gene for efficient mutant selection are described.

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: The disclosure describes highly specific tryptase polyclonal antibodies, and a method to purify the antibodies. Specifically, the invention relates to polyclonal antibodies which have the capacity to capture tryptase out of solution, a process to generate the antibodies, and an enzyme-linked immunosorbent assay (ELISA) for human tryptase which utilizes the antibodies.

Abstract: The present invention relates to single and dual-reporter luminescence assays utilizing general and specific reagents to quench enzyme-mediated reactions. In one embodiment of the invention, a reagent is added to the assay which non-specifically quenches enzyme-mediated luminescent reactions. In another embodiment of the invention, a reagent is added to the assay which simultaneously quenches one enzyme-mediated luminescent reaction while activating another distinct enzyme-mediated luminescent reaction. An assay kit containing specific quench reagents, and the reagents themselves are also disclosed.

Abstract: The present invention provides novel dyes for staining proteins, particularly those in electrophoretic gels, and detecting the proteins with high sensitivity. The dyes of the invention are derivatives of known dyes that are more hydrophobic than the corresponding, known dyes and, in electrophoretic gels, including polyacrylamide gels with sodium dodecyl sulfate, form easily detectable, non-covalent complexes with proteins. In some cases, the corresponding, known dyes bind to protein covalently. The complexes of protein with some dyes of the invention can be detected with high sensitivity simply by observation of the color of the complexes by eye. The complexes of protein with other dyes of the invention are detected by observation of the fluorescence of the complexes. Dyes of the invention include derivatives of Coomassie.RTM. dyes, acid blue 25, dansyl chloride, fluorescein, 2-methyl-2,4-diphenyl-3(2H)-furanone, and (7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino alkanoates.

Abstract: Processes are provided for removal of cells as cell pellets from liquid milk samples, or from cultures or extracts of other food materials or other materials of biological origin. The concentrated cells in the pellet can be analyzed by various techniques to determine the relative cell count, such as by lysing of the cells followed by measurement of ATP. Nucleic amplification, as by the polymerase chain reaction method, can be carried out using the cellular pellet directly, without need for isolation of nucleic acid from the cells. After the amplification, an assay can be carried out for amplified nucleic acid segment indicative of the presence of cells of interest in the sample. The invention thus provides methods for obtaining cellular components from samples of milk, and cultures or extracts of other materials, including food materials, and for determining relative contamination of milk and such other materials by microorganisms. The invention also provides kits for carrying out its various methods.