Abstract: The present application relates to mutated RNA polymerases from bacteriophages that have increased stability, for example under high temperature conditions. One example of bacteriophage encoded RNA polymerase is the T7 RNA polymerase. T7 is a bacteriophage capable of infecting E. coli cells. Examples of other E. coli infecting T7-like bacteriophages are T3, øI, øII, W31, H, Y, A1, croC21, C22 and C23. An example of a Salmonella typhimurium infecting bacteriophage is SP6. The present invention is concerned with the RNA polymerases of T7-like bacteriophages that have been mutated. Due to these mutations the RNAP's have an increased stability. Preferred mutated RNA polymerases according to the invention are mutant RNA polymerases from T7 or SP3 bacteriophages. Due to the high homology between these enzymes, mutations in the T7 gene 1 sequence are likely to have the same effect in the corresponding gene sequence of the T3 bacteriophage.

Abstract: The invention relates to a novel mutant of tissue factor pathway inhibitor (TFPI) protein and its corresponding DNA sequence. The mutant can be found in humans who show or may show an increased risk of thrombotic diseases. By screening samples of human blood for said DNA or fragments of it, it is possible to predict a disposition of thrombotic disorders by which prophylactic application or measures can be initiated.

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 ERCC1 expression levels in fixed or fixed and paraffin embedded tissues and prognosticate the probable resistance of a patient's tumor to treatment with platinum-based therapies by examination of the amount of ERCC1 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 ERCC1 and methods comprising their use for detecting levels of ERCC1 mRNA.

Abstract: Method for analyzing a phyletic lineage of scallop, which comprises the steps of sequencing a mitochondrial DNA of the scallop to determine a nucleotide base sequence of the mitochondrial DNA, wherein the nucleotide base sequence includes a non-coding region containing a nucleotide base substitution locus which shows a sequence polymorphism indicative of a particular lineage of the scallop. Mitochondrial DNA of the scallop is amplified by PCR, using a suitable primer set designed on the basis of nucleotide base sequences conserved among known shellfish mitochondrial 16S rRNA and 12S rRNA genes, followed by sequencing to determine a non-coding region therein. In such non-coding region, a nucleotide base substitution locus is located, whereby a particular lineage of the scallop is determined. Based on those steps, a Japanese scallop, Patinopecten yessoensis, is analyzed as to the nucleotide base sequence and lineage thereof.

Abstract: A method for discriminating between two or more alleles which differ by only a single nucleotide is provided. This method is suitable for use in solution or on a solid phase and employs the use of restriction enzyme digestion of DNA in combination with Melting Curve Analysis. Also contemplated are specific markers to be included in melting curve analysis and melting curve genotyping experiments.

Abstract: The present invention provides a method for the rapid simultaneous production of a plurality of single-stranded DNA circles having a predetermined size and nucleotide sequence using pre-designed hairpin oligonucleotides containing complementary sequences for directing ligation to form dumbbell-shaped monomers followed by heat denaturation to yield single-stranded DNA circles.

Abstract: This invention relates to an isolated nucleic acid fragment encoding a sterol metabolism enzyme. The invention also relates to the construction of a chimeric gene encoding all or a portion of the sterol metabolism enzyme, in sense or antisense orientation, wherein expression of the chimeric gene results in production of altered levels of the sterol metabolism enzyme in a transformed host cell.

Abstract: The present invention provides a high throughput method for genotyping microsatellite markers. The technology uses a combination of oligonucleotides in a selective ligation reaction that discriminates mono-, di-, tri-, tetra-, penta-, hexa-, hepta-, octa- and nona-nucleotide repeated alleles in amplified DNA from individuals.

Abstract: The present invention provides a method for exploring low molecular weight compounds which regulate positively or negatively the expression of the human BMP-2 with reference to a reporter activity by using 5′ upstream region gene containing the human BMP-2 promoter and an animal cell introduced with a recombinant expression vector which has been connected to an appropriate reporter gene. The low molecular weight susbtances and their derivatives obtained by the present method have morphogenetic activity and inhibiting activity for bone and cartilage through the expression of human BMP-2 and are useful as preventive or therapeutic agents for bone and cartilage diseases.

Abstract: An object of the present invention is to establish a method for amplifying an RNA in a biologically-derived sample by preparing a reaction solution in which a nucleic acid synthesis is not inhibited even in the presence of various biologically-derived impurities and as well as to enable a convenient, rapid and highly sensitive analysis of the RNA in the sample. In a method of the present invention, a reaction solution having a pH higher than that of an ordinarily employed reaction solution and/or containing a polyamine and/or a sulfated polysaccharide.

Abstract: Oligonucleotides are provided which are useful as primers to initiate the amplification of a segment of Histoplasma capsulatum DNA that is specific to H. capsulatum, using the polymerase chain reaction. A method of detecting the presence of H. capsulatum in a sample using a nested, or two-stage, PCR assay is also provided. The outer pair of primers, for use in the first stage of the assay, are fungal-specific oligonucleotides; the inner pair of primers, for use in the second stage, are oligonucleotides specific for H. capsulatum.

Abstract: This invention provides a method of sorting genes comprising: (1) preparing ds cDNA molecules from mRNA molecules (2) digesting the ds cDNA molecules (3) ligating to the digested cDNA molecules a set of dsDNA oligonucleotide adaptors; (4) amplifying the ligated cDNA molecules; and (5) sorting the amplified cDNA molecules into nonoverlapping groups. This invention also provides two additional methods of sorting genes. This invention further provides a method of making sub-libraries of ligation sets.

Abstract: The present invention relates to the detection of the presence of reverse transcriptase enzymatic activity in eukaryotic cell lines. Specifically, the present invention comprises a method for discriminating between endogenous reverse transcriptase activity in a substrate cell line (e.g., avian) used as substrates for the manufacture of biological products (including viral vaccines) from exogenous infectious retrovirus by using a combination of transmissibility assay (viral replication) and PBRT assay (RT-PCR). Appropriate levels of viral amplification and number of PCR cycles are determined and employed that permit detection of exogenous infectious RT activity but not endogenous RT activity.

Abstract: This invention relates to prevention and/or treatment of antibiotic associated diarrhea, including Clostridium difficile associated diarrhea (CDAD), pseudomembranous colitis (PMC) and other conditions associated with C. difficile infection, using oligosaccharide compositions which bind C. difficile toxin B. More specifically, the invention concerns neutralization of C. difficile toxin B associated with such conditions.

Abstract: The present invention provides methods of determining relative copy number of target nucleic acid sequences and precise mapping of chromosomal abnormalities associated with disease. The methods of the invention use target nucleic acid sequences immobilized on a solid surface, to which a sample comprising two sets of differentially labeled nucleic acid sequences are hybridized. The hybridization of the labeled nucleic acid sequences to the solid surface is then detected using standard techniques.

Abstract: The present invention provides a method for exploring low molecular weight compounds which regulate positively or negatively the expression of human BMP-4 with reference to a reporter activity by using 5′ upstream region gene containing the human BMP-4 promoter and an animal cell introduced with a recombinant expression vector that has been connected to an appropriate reporter gene. The low molecular weight compounds and their derivatives obtained by the present method have morphogenetic activity and inhibiting activity for bone and cartilage through the expression of human BMP-4 and are useful as preventive or therapeutic agents for cartilage and bone diseases.

Abstract: The present invention provides isolated an polynucleotide comprising a polymorphic nucleotide sequence from a diacylglycerol acyltransferase (DGAT) gene, wherein at least one a polymorphism is associated with a condition relating to DGAT activity. The invention further provides diagnostic assays for detecting the presence in a nucleic acid sample of a polymorphism in a DGAT gene that is associated with a condition relating to DGAT activity, and for detecting a propensity to develop a condition associated with DGAT activity. The invention further provides treatment methods.

Abstract: The present invention provides methods of using miniature inverted repeat transposable elements, including producing a DNA fingerprint of an individual, detecting at least one polymorphism between the nucleic acid fragments of two individuals, and correlating the presence of an amplified fragment to a phenotype.

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: The invention concerns a device for collection of magnetic particles. The invention further concerns a system for transfer of material from a plurality of source vessels to a plurality of target vessels, the device comprising a plurality of collecting members each of which can be manipulated independently. The invention further concerns a method for detection of biological entities being fluorescent label, wherein the entities are bound to magnetic particles. The fluorescent emission is concentrated by clustering the magnetic particles utilizing magnetic force.