Abstract: A buffer composition, method and kit for hybridizing microarrays of nucleic acids bound to an adsorbed polymer surface of a siliceous substrate provide an envelope of conditions to hybridize nucleic acid targets, while preserving theintactness of the adsorbed polymer surface of the array. The buffer composition comprises a non-chelating buffering agent, a pH within a range of pH 6.4 and 7.5, a monovalent cation having a monovalent cation concentration that ranges from about 0.01 M to about 2.0 M, and optionally relatively lower concentrations of a chelating agent and an ionic surfactant. The total cation concentration of the buffer composition ranges from about 0.02 M to about 2.0 M. The method comprises incubating the targets with the microarray in the buffer composition at a temperature between about 55° C. and 70° C.

Abstract: The invention provides a method for reducing background in hybridization reactions of nucleic acids involving at least two homologous probes, wherein at least one of the probes is non-linear, or two homologous target sequences and a non-linear probe. Background is reduced by introducing an intended mismatch with a target sequence in at least one of the probes. The presence of the mismatch reduces the specificity of probes not entirely complementary to a target sequence to such an extent that the background signal is reduced. A set of mixed homologous probes, wherein at least one of the probes is non-linear, comprising such specific mismatch is also provided. The set can be used for the detection of variants of a family of nucleic acids, for instance a number of HIV variants. The invention also provides kits for carrying out the methods according to the invention.

Abstract: The invention concerns mass spectrometric analysis of known mutation sites in the genome, such as single nucleotide polymorphisms (SNPs). The invention uses minor amounts of primers with photocleavable linkers, intermixed with a major amounts of primers without linkers, to produce short mutation-containing DNA sequences by enzymatic amplification procedures such as polymerase chain reactions (PCR). After this single amplification procedure, the linker-containing PCR by-products are extracted, washed and photolytically cleaved. Short oligonucleotides are produced which facilitate mass spectrometric analysis. Additionally to the use of linkers, some types of primers may contain blockers which stop the polymerase copying process to achieve even shorter oligonucleotides for analysis.

Abstract: Methods of end-labeling ribonucleic acids with non-radioactively labeled ribonucleotides, and particularly fluorescently labeled ribonucleotides, are provided. In the subject methods, a ribonucleic acid is contacted with a non-radioactively labeled ribonucleotide in the presence of a prokaryotic, usually bacterial, poly(A) polymerase under conditions sufficient for covalent bonding of the labeled ribonucleotide to the 3′ end of the ribonucleic acid to occur. Also provided are kits for practicing the subject method. The subject methods and kits find use in a variety of applications where labeling of the 3′ end of a ribonucleic acid with a non-radioactive label, particularly a fluorescent label, is desired.

Abstract: The present invention provides a method for separation of a particulate matrix from a solution while reducing loss of particles during separation steps. Methods are also disclosed for isolation of molecules of interest using affinity particles or beads, wherein at least one step of the isolation is conducted in the presence of a detergent. The presence of detergent reduces the loss of matrix particles and enhances reproducibility and yield of the molecule of interest. The invention makes possible the manual and automated processing of affinity beads, especially magnetic beads, in multi-well as well as single-well vessels with significantly reduced bead loss, as compared with similar processes conducted in the absence of detergent.

Abstract: The present invention describes polynucleotides and polypeptides associated with PPAR&agr;-mediated pathways that are useful as therapeutic compositions in method for the treatment of peroxisomal disorders. These polynucleotides and polypeptides were identified through the use of differentioal gene expression analysis. In particular, the present invention discloses eleven novel gene fragments, and numerous single nucleotide polymorphisms, located in previously disclosed genes, all of which have been discovered to be associated with PPAR&agr;-mediated pathways.

Abstract: Fluorescent labels having at least one donor and at least one acceptor fluorophore bonded to a polymeric backbone in energy transfer relationship, as well as methods for their use, are provided. Of particular interest are the subject labels wherein the polymeric backbone is a nucleic acid and the donor fluorophore is bonded to the 5′ terminus of said nucleic acid. Such labels find use as primers in applications involving nucleic acid chain extension, such as sequencing, PCR and the like.

Abstract: The present invention relates to genetic mutations in mitochondrial genes that segregate with diabetes mellitus. The invention provides methods for detecting such mutations, as a diagnostic for diabetes mellitus, either before or after the onset of clinical symptoms. Examples of specific mutations in the ATP synthase 8/6 sequence and tRNALys sequence are given. The invention also provides treatments for dysfunctions due to genes for mitochondrial functions that segregate with diabetes mellitus. Cybrid cell lines are described which are useful as model systems for the study of the mitochondrial metabolic disorders that are associated with diabetes mellitus, and for identifying therapeutic compounds and treatments for this disease.

Abstract: This specification relates to the field of molecular biology and provides novel methods and reagents for preserving and protecting the ribonucleic acid (RNA) content of samples from degradation prior to RNA isolation. This preservation may be accomplished without ultra-low temperature storage or disruption of the tissue.

Abstract: The invention relates to a complex of specifically complex-forming proteins which are not naturally occurring, comprising the following components: a) at least one ligand specific for a target structure, b) at least one protein comprising a mutated dimerization domain, the mutated dimerization domain having been derived by mutation of a naturally occurring dimerization domain, it being possible for this mutated dimerization domain to interact specifically with component c) and the component b) being connected covalently to the component a), c) at least one protein comprising a mutated dimerization domain, the mutated dimerization domain having been derived by mutation of a naturally occurring dimerization domain, it being possible for this mutated dimerization domain to interact specifically with component b) and the component c) is linked covalently to the component d), and d) at least one effector.

Abstract: A process is provided for labeling with signal amplification a ribonucleic acid (RNA), comprising fragmenting the RNA to form RNA fragments, fixing a first ligand to a terminal phosphate located at least one of the 3′ end and the 5′ end of each of a plurality of the RNA fragments, the terminal phosphate having been released during the fragmentation, and binding a plurality of labeling agents to the first ligand on each of a plurality of the fragments.

Abstract: The present invention relates to a method of detecting nucleic acids and particularly to a method of analyzing for the presence and/or amount of a nucleic acid.

Abstract: Novel polynucleotides isolated from Lactobacillus rhamnosus, as well as probes and primers, genetic constructs comprising the polynucleotides, biological materials, including plants, microorganisms and multicellular organisms incorporating the polynucleotides, polypeptides expressed by the polynucleotides, and methods for using the polynucleotides and polypeptides are disclosed.

Abstract: The conventional synthesis of nucleic acid polymers from several oligonucleotides comprises several cycles of intermediate product synthesis, purification of the intermediate products and synthesis of a full length product (up to a maximum length of approximately 1000 nucleotides). The novel method shall be carried out in one reaction step and result in nucleic acid polymers of more than 1000 nucleotides. According to the invention, two or more linkable oligonucleotides are provided that in a continuous arrangement and after linkage can form a primary strand, and one or more non-linkable oligonucleotides are provided, each of the non-linkable oligonucleotides comprising two adjoining regions, the first of which is complementary to the 3′ end of a linkable oligonucleotide and the second of which is complementary to the 5′ end of a further linkable oligonucleotide.

Abstract: The present invention relates to a polynucleotide in isolated form, which polynucleotide codes for a protein with the activity of the enzyme L-galactono-&ggr;-lactone dehydrogenase, which polynucleotide comprises at least the L-galactono-&ggr;-lactone dehydrogenase activity-determining parts of the coding part of the nucleotide sequence or a sequence derived therefrom on the basis of the degeneration of the genetic code. The invention further relates to the use of the polynucleotide in the production of transgenic plants, plant cells, or other eukaryotic cells.

Abstract: Novel and improved processes for isolating organophosphorus hydrolase enzyme from an aqueous solution and obtaining substantially purified enzyme at high yield are provided, as well as compositions, including storage stable lyophilyzed organophosphorus hydrolase enzyme compositions, that are prepared by the provided methods. The organophosphorus hydrolase enzyme is purified by contacting an aqueous solution of cell free bacterial proteins with a strong cation exchange resin, the aqueous solution comprising soluble organophosphorus hydrolase enzyme, washing the strong cation exchange resin with a washing buffer to remove unbound proteins from the strong cation exchange resin, eluting proteins that remain bound to the strong cation exchange resin by washing the resin with an eluting buffer comprising salt in a concentration that starts at about zero and is raised during the eluting process to about 0.5M, and detecting and collecting eluate comprising a protein having organophosphorus hydrolase enzyme activity.

Abstract: A set of contiguous and partially overlapping cDNA sequences and polypeptides encoded thereby, designated as PCIGF and transcribed from prostate tissue, is described. These sequences are useful for the detecting, diagnosing, staging, monitoring, prognosticating, in vivo imaging, preventing or treating, or determining the predisposition of an individual to diseases and conditions of the prostate, such as prostate cancer. Also provided are antibodies which specifically bind to PCIGF-encoded polypeptide or protein, and agonists or inhibitors which prevent action of the tissue-specific PCIGF polypeptide, which molecules are useful for the therapeutic treatment of prostate diseases, tumors or metastases.

Abstract: The present invention relates to an oligonucleotide having a novel structure and a method of synthesizing nucleic acid by using the same as a primer. This oligonucleotide is provided at the 5′-side of the primer with a nucleotide sequence substantially the same as a region synthesized with this primer as the origin of synthesis. The present invention realizes synthesis of nucleic acid based on an isothermal reaction with a simple constitution of reagents. Further, the present invention provides a method of synthesizing highly specific nucleic acid on the basis of this method of synthesizing nucleic acid.

Abstract: A method of stabilizing a useful protein by mixing a useful protein and an aqueous solution of a compound having the basic structure of arabic acid, and a stabilizied useful protein compositon containing a useful protein and a compound having the basic structure of arabic acid; gum arabic is preferred as a compound having the basic structure of arabic acid, and examples of a useful protein include cytokine and interferon; and a production method for canine interferon-&ggr; and stabilization thereof.

Abstract: Signal primers are employed for detection of nucleic acid target sequences by fluorescence quenching mechanisms. The signal primer comprises a first and a second oligonucleotide and is partially single-stranded and partially double-stranded. In the presence of target, the second oligonucleotide of the signal primer is displaced from the first and a conformational change in a reporter probe occurs which changes the distance between the members of a donor/quencher dye pair linked to the reporter probe. The change in proximity between the dyes causes an increase or a decrease in fluorescence quenching, which is detected as an indication of the presence of the target sequence.