Abstract: A process for synthesizing an oligonucleotide comprising linking a nucleoside phosphate to a solid support, through the heterocyclic moiety of the nucleoside, coupling a mono- or oligonucleotide to the nucleoside phosphate through its phosphate moiety, in at least one step enzymatically lengthening the mono- or oligonucleotide, cleaving the resultant oligonucleotide from the solid support-nucleoside phosphate at the phosphate moiety of the nucleoside, and separating the oligonucleotide. After cleaving and separating the solid support-nucleoside phosphate is recycled for further coupling. Advantageously the solid support-nucleoside phosphate is phosphorylated between separation and recycling.

Abstract: This invention concerns a coordination complex and salts and optically resolved enantiomers thereof, of the formula (R).sub.3 --M, wherein R comprises 1,10-phenanthroline or a substituted derivative thereof, M comprises a suitable transition metal, e.g. ruthenium(II) or cobalt(III), and R is bonded by M by a coordination bond.The complexes of this invention are useful in methods for labeling, nicking and cleaving DNA. The lambda enantiomer of complexes of this invention is useful in methods for specifically labeling, detecting, nicking and cleaving Z-DNA.The complexes may also be used in a method for killing tumor cells and may be combined with a pharmaceutically acceptable carrier to form a pharmaceutical composition for the treatment of tumor cells in a subject. The invention further concerns methods for treating a subject afflicted with tumor cells.

Abstract: Nonradiometric polynucleotide probe compositions comprising zymogen-activating polypeptide reporter moieties are disclosed, in addition to detection methods and systems employing said probes.

Abstract: The rate of hybridization between two complementary polynucleotide segments in an aqueous medium is increased by the presence of the anionic polymers polyacrylate and polymethacrylate. The acceleration effect is particularly useful in nucleic acid hybridization assays involving immobilization of sample nucleic acids and the use of labeled probes. Nonspecific binding of probe to nitrocellulose supports is substantially lower in the presence of the present polymers than in the presence of the prior art accelerator dextran sulfate. Polyacrylate is particularly advantageous since it has been found to be effective at low concentrations and is significantly less expensive than the prior art compound.

Abstract: Method of detecting the presence of Salmonella in a food sample including providing at least one DNA probe which is capable of selectively hybridizing to Salmonella DNA to form detectable complexes, contacting the DNA probes with the bacteria in the food sample under conditions which allow the probe to hybridize to Salmonella DNA present in the food sample to form hybrid DNA complexes, and detecting the hybrid DNA complexes as an indication of the presence of Salmonella in the food sample.

Abstract: A method for identifying and isolating clones containing DNA coding for a desired protein is described. DNA prepared from a cell that expresses the desired protein is inserted into an isolation expression vector having means for replication (as a means of producing DNA) and a suitable promoter for expression of said DNA in a predetermined mammalian host cell as well as means for replication in a bacterial cell. The transient expression vector is then inserted into a bacterial cell for replication of the DNA. Pools of DNA, prepared from a predetermined number of bacterial clones so that the nucleic acids (DNA and RNA) is substantially free of other bacterial contaminants are transfected or microinjected into mammalian host cells and conditioned medium from growing such cells is tested for the presence of the desired protein. Positive pools are selected and the clones used to make the pool are screened to identify and isolate the clone containing the desired DNA.

Abstract: In a method for determining a particular polynucleotide sequence in a test medium containing single stranded nucleic acids wherein the sample is subjected to a hybridization reaction with a labeled detection probe having a substantially complementary polynucleotide sequence, and wherein after hybridization the label in said probe is assayed, the improvement wherein the label in said labeled probe comprises a fluorescent nucleotide which is linked by a phosphate ester linkage to said probe. Probes and kits therefor are also provided.

Abstract: A method for detecting the presence of the gene for Huntington's disease in a subject which comprises analyzing the human chromosome 4 of the subject for a DNA polymorphism, preferably a restriction fragment length polymorphism (RFLP), linked to Huntington's Disease.

Abstract: Mouse monoclonal antibodies to several cell antigens of human ovarian, cervical and endometrial carcinomas have been produced and characterized. The distribution of the antigens was determined by mixed hemagglutination assays on 153 normal and malignant cell cultures of various types, and by immunoperoxidase staining of frozen sections of 27 normal adult and 24 fetal tissues. five monoclonal antibodies representative of five classes of mAb raised to restricted ovarian, cervical and endometrial cells were tested extensively producing mAb reactive with cancer but not normal cells. One such mAb, MF116 was readily detected in the spent culture medium of metabolically radiolabeled cells. These antibodies, reacting with relatively restricted cell surface antigens, are useful in the analysis of epithelial cell differentiation, in cancer diagnosis and therapy and in tissue typing of normal or abnormal cells.

Abstract: Dyes are provided comprising phycobiliproteins having covalently coupled thereto a plurality of small molecular weight organic dyes under conditions permitting energy transfer mechanisms to operate therebetween. The resultant label may then be used in virtually any immunoassay format.

Abstract: Self-sufficient incubation assembly for the in vitro cultivation of microorganisms such as bacteria and for being energized by a self-contained energy source, including a heater for heating means, such as a culture growth dish assembly or a cuvette, for receiving a culture growth medium seeded with microorganisms, to a physiological temperature to cultivate the microorganisms, and electrical circuitry which interconnects the heater with the energy source and which includes a temperature control element in intimate physical contact with the seeded culture growth receiving means to cause the temperature of the control element to be substantially the same as the temperature of the medium; the electrical circuit in operation produces heat and due to its intimate physical contact with the means for receiving the seeded culture growth medium supplements the heating of the medium by the heater and the supplementation reduces the total energy required to be supplied by the energy source to cultivate the microorganism

Abstract: Compositions and methods for potentiating the cytotoxic activity of immunogotoxin conjugates are provided. The compositions of the present invention include a selective binding agent such as an antibody coupled to a toxin B chain moiety such as ricin B chain.

Abstract: A device is described which provides a totally enclosed and secure environment for the culture of cells and for the production of biologicals, pharmaceuticals, and other cell-derived products of commercial value. The device comprises several modules of differing functions: regulating the cellular growth environment, providing a suitable cell growth substrate or environment, or separating of desired product form interfering substances. Each module comprises a series of membranes separated by a solid support material which is channeled to provide a series of parallel capillaries for the flow of nutrients, environmental regulating fluids, or gas exchange. The structure and composition of the separating membranes limits the nature, rate, and size of the particular material exchanged across an individual membrane into or from an adjacent compartment. The capillaries are so dimensioned that the nutrients and gases are diffused over distances typical of human tissue.

Abstract: A method of detecting a mutation of a specific nucleotide base in a target nucleic acid chain comprises: (a) hybridizing a labelled probe to the target to form a hybrid in which one end of the probe is positioned adjacent the specific base; (b) adding a nucleotide derivative, e.g. a thionucleotide, under conditions to cause it to join to the end of the probe if it is complementary to the specific base; (c) digesting the hybrid using an exonuclease enzyme under conditions such that the nucleotide derivative protects the probe from digestion; and observing the presence or absence of label attached to the target. The method can be used to detect mutations even when these are not present at restriction enzyme cleavage sites, and does not require the preliminary steps of restriction digestion, gel electrophoresis and DNA (or RNA) blotting.

Abstract: Methods for the detection and identification of unknown pathogens or genetic entities from samples suspected of containing the same involve the use of novel wicking techniques whereby sample DNA/RNA becomes readily available on a porous, inert, positively charged support in single stranded form for hybridization with a hybridization probe having a nucleotide sequence substantially complementary to a nucleotide sequence of the sample DNA/RNA. Such methods permit the detection and identification of unknown pathogens and genetic entities in a shorter period of time than that required by conventional methods. Lysis solutions for rapidly lysing bacterial cells without the aid of enzymes are also disclosed.

Abstract: Assays are provided employing particles and absorbent particles, wherein the absorbent particles substantially inhibit fluorescence when bound to the fluorescent particles through specific non-covalent binding.

Abstract: A reagent and merchandizing kit for use in the diagnosis of syphilis by hemagglutination of an anitgen obtained from a culture of pathogenic Treponema pallidum Nichols and which is sensitized on carrier particles in the presence of antibody wherein said antigen is substantially devoid of proteinic fractions of said culture having a specific gravity of less than 1.01, and methods of preparing the same.

Abstract: The present invention provides a stable, continuous human myeloma cell line which is capable of hybridization with antibody-producing cells of humans and other animals, the cell line being a mutant of GM 1500 human B cells and being deficient in hypoxanthine phosphoribosyltransferase. The invention also comprises processes for the production of hybrid cells employing the stable, HPRT-deficient human myeloma cell line, and processes for the production of antibodies employing such hybrid cells.

Abstract: An agglutination assay reagent and method. The reagent is composed of liposomes predominantly in the 1 to 20 micron diameter size range. Each liposome has a surface array of laterally mobile ligand molecules, at a surface concentration adapted to produce reagent agglutination within about 5 minutes, when the reagent is incubated at room temperature with a multivalent ligand-binding analyte. A dye in the liposomes allows such agglutination to be visualized easily without magnification.

Abstract: A microprocessor controlled, general purpose gene synthesizer for programmably synthesizing selected nucleotide sequences. Depending upon a programmed chemistry, metered volumes of selected bases and reagent/solvents are sequentially metered into the bottom of a reaction cell containing solid-supported nucleotides, where desired linkages are achieved in an agitated suspension. Self-refilling syringe metering and electrically initiated, pneumatically operated valving insures economical, non-contaminated synthesis. Preprogrammed, selectable chemistries and operator programmed sequences are operatively coupled under microprocessor control to ensure a general purpose synthesizer.