Abstract: Carboranyl-containing nucleosides and oligonucleotides are provided for use in boron neutron capture therapy (BNCT) and for other therapeutic and diagnostic purposes.

Abstract: The present invention provides oligomers which are specifically hybridizable with a selected sequence of RNA or DNA wherein at least one of the nucleoside moieties of the oligomer is modified to include an aminooxy linkage. These oligomers are useful for diagnostic, therapeutic and investigative purposes.

Abstract: Methods for testing oligonucleotide arrays are disclosed including methods for testing the efficiency of nucleotide coupling; methods for testing amounts of deprotected oligonucleotides; methods for determining amounts of depurinated oligonucleotides; and methods of detecting the presence of cleavable structural features, such as double-stranded nucleic acids.

Abstract: This invention provides reagents, libraries and sets of the reagents, and assay methods using the reagents, the reagents comprising an analyte moiety and a tag moiety, wherein the tag moiety contains information defining the identify and location of the analyte residues of the analyte moiety which is detectable by mass spectrometry.

Abstract: A novel method for the labeling of oligonucleotides which results in the economical synthesis of 5′ labeled molecules. A set of suitably protected and carefully selected set of amino linkers, a modified deprotination/cleavage protocol and standard coupling methodologies to are used to allow for the convergent synthesis of any number of labeled oligonucleotides.

Abstract: The invention is directed to synthetic receptor(s) which comprises a polyfunctional organic template covalently linked to two or more oligomers which may independently be the same or different and may independently be straight chain, cyclic or branched. The template may be linked to an identifier which uniquely defines the synthetic receptor. The identifier is a stable chemical molecule or a plurality of stable chemical molecules distinguishable and detectable to picomolar levels or may be an oligonucleotide. In a preferred embodiment, the template is covalently linked to a solid support which is linked to an identifier. In addition, the invention includes methods of preparing synthetic receptors and synthetic receptor libraries. The synthetic library may be linked with identifiers such that the library comprises a plurality of different synthetic receptor members.

Abstract: Nucleoside-5′-triphosphates and phosphoramidites which carry a residue absorbing in the long wavelength region, preferably a carbocyanine group of the general formula (I), on the base portion or on the phosphorus atom in which R1 and R2 each denote hydrogen or together form a phenyl residue; R3 denotes hydrogen if linkage with the nucleotide is via the R4 position or it denotes a —NHCS— group if linkage with the nucleotide is via the R3 position; both R4 and R5, or R5, alone denote an alkylsulfonyl group with n being a number from 3 to 5 or R4 represents a —NHCS— group with n being a number from 3 to 8, as well as the use of the compounds to label, detect and sequence nucleic acids.

Abstract: A screening method which applies the principle of synthetic lethality to a gene of interest in non-yeast eukaryotic cells. The method may be used to screen either a chemical library in order to identify a molecule having a gene-specific lethal property in the cells, or to screen a group of DNA molecules in order to identify among them one or more modulators of gene function which are synergistically lethal to the cells together with an incapacitated gene of interest. Also described are episomal survival plasmids and kits which may be used with the method.

Abstract: Novel compounds are provided which are useful as linking groups in chemical synthesis, preferably in the solid phase synthesis of oligonucleotides and polypeptides. These compounds are generally photolabile and comprise protecting groups which can be removed by photolysis to unmask a reactive group. The protecting group has the general formula Ar—C(R1)(R2)—O—C(O)— wherein: Ar is an optionally substituted fused polycyclic aryl or heteroaromatic group or a vinylogous derivative thereof; R1 and R2 are independently H, optionally substituted alkyl, alkenyl or alkynyl, optionally substituted aryl or optionally substituted heteroaromatic, or a vinylogous derivative of the foregoing; and X is a leaving group, a chemical fragment linked to Ar—C(R1)(R2)—O—C(O)— via a heteroatom, or a solid support; provided that when Ar is 1-pyrenyl and R1 and R2 are H, X is not linked to Ar—C(R1)(R2)—O—C(O)— via a nitrogen atom.

Abstract: A method for fluorophore bias removal in microarray experiments in which the fluorophores used in microarray experiment pairs are reversed. Further, a method for calculating the individual errors associated with each measurement made in nominally repeated microarray experiments. This error measurement is optionally coupled with rank based methods in order to determine a probability that a cellular constituent is up or down regulated in response to a perturbation. Finally, a method for determining the confidence in the weighted average of the expression level of a cellular constituent in nominally repeated microarray experiments.

Abstract: An oligonucleotide for detection or amplification of a gene selected from the group consisting of Vibrio parahaemolyticus thermostable direct hemolysin-related hemolysin genes (trh1 and trh2) and Vibrio parahaemolyticus thermostable direct hemolysin gene (tdh2) or RNA derived therefrom is provided. Further, method for detecting trh1, trh2 or tdh2 using said oligonucleotide is provided.

Abstract: The present invention relates to fluorescent probes which can be used in multicolor fluorescence in situ hybridization, and mainly chromosome painting. The probes intended for labeling a chromosome are such that they are composed of a set of DNA segments which are more represented in certain chromosome bands and which are obtained by IRS-PCR amplification from said chromosomes using PCR primers specific for the repeated and dispersed Alu and LINE DNA sequences. The invention comprises, in addition, methods of producing said probes, multicolor FISH methods which can use said probes as well as diagnostic kits comprising them. Finally, the invention comprises combinations of fluorophores and optical filters.

Abstract: Disclosed is a method of detecting specific nucleic acids using an oligonucleotide linked to a cleavable tag. The presence of a specific nucleic acid in a population of nucleic acids is determined by hybridizing an oligonucleotide containing the tag to a population of nucleic acids, separating hybridizing bound oligonucleotides, and then removing and identifying the tag. Also provided are compositions and kits comprising oligonucleotides linked to a cleavable tag.

Abstract: A method is disclosed for detecting the presence of a difference between two related nucleic acid sequences. In the method a complex is formed comprising both strands of each sequence. Each member of at least one pair of non-complementary strands within the complex have labels. The association of the labels as part of the complex is determined as an indication of the presence of a difference between the two related sequences. The complex generally comprises a Holliday junction. In one aspect a medium suspected of containing said two related nucleic acid sequences is treated to provide partial duplexes having non-complementary tailed portions at one end. The double stranded portions of the partial duplexes are identical except for said difference.

Abstract: Disclosed is a method of detecting specific nucleic acids using an oligonucleotide linked to a cleavable tag. The presence of a specific nucleic acid in a population of nucleic acids is determined by hybridizing an oligonucleotide containing the tag to a population of nucleic acids, separating hybridizing bound oligonucleotides, and then removing and identifying the tag. Also provided are compositions and kits comprising oligonucleotides linked to a cleavable tag.

Abstract: A multilayered microfluidic DNA analysis system includes a cell lysis chamber, a DNA separation chamber, a DNA amplification chamber, and a DNA detection system. The multilayered microfluidic DNA analysis system is provided as a substantially monolithic structure formed from a plurality of green-sheet layers sintered together. The substantially monolithic structure has defined therein a means for heating the DNA amplification chamber and a means for cooling the DNA amplification chamber. The means for heating and means for cooling operate to cycle the temperature of the DNA amplification chamber as required for performing a DNA amplification process, such as PCR.

Abstract: A labeling reagent for use in oligonucleotide (“oligo”) synthesis, as well as a method of preparing such a labeling reagent, a method of using such a reagent for synthesizing a labeled oligonucleotide, and an oligonucleotide prepared using such a reagent. The reagent can be used to label either the 3′ or 5′ termini of a synthesized oligo, and/or for one or more positions along the oligo. The labeling reagent can be prepared by a reaction scheme that involves the initial preparation of hydroxyacids, tritylated hydroxyacids and coupling of such derivatives to diamine, wherein the amine function serves at an attachment point for labels and the hydroxyl groups can either be used to immoblize the molecules to support or can be converted to provide a phosphorylating reagent.

Abstract: The present invention provides a means to identify the alleles present in a DNA-containing sample by providing subsets of loci for amplification by multiplex PCR. The loci include the thirteen CODIS short tandem repeat (STR) loci and amelogenin. The loci within each subset are grouped so that, upon PCR amplification, the amplicons produced within a given subset do not overlap. Differential labeling of subsets makes it possible to further group the subsets into compound multiplexes for co-amplification in a single reaction vessel, and analysis in a single electrophoretic channel.

Abstract: Compounds are described comprising purines substituted at the C-8 position and pyrimidines substituted at the C-4 position with a linker and quencher. These compounds, when incorporated into hairpin oligonucleotides quench the fluorescence of fluorophores linked to the 5′-terminus of the oligonucleotide. These nucleotide-quencher compounds are also easily incorporated into oligonucleotides using conventional or automated oligonucleotide synthetic techniques.

Abstract: Provided is a polynucleotide comprising mRNA, rRNA or viral RNA, comprising ribose rings that are covalently modified at the 2′-OH position. Further provided are methods for producing a double-stranded oligo- or polynucleotide from a template comprising an oligo- or polyribonucleotide, a proportion of the ribose rings of which are covalently modified at the 2′-OH position to bear a substituent which enables replication of the template by the nucleic acid polymerase. Also provided is use of a poly-nucleotide comprising mRNA, rRNA or viral RNA, a proportion of the ribose rings of which are covalently modified at the 2′-OH position, in a hybridization reaction.

Abstract: Methods and compositions to label oligonucleotides and analogs directly on a solid-support having the structure where S is a solid-support, A is a cleavable linker, X is a moiety with three or more attachment sites, L is a label, Y is a nucleophile, i.e. O, NH, NR or S, and P1 is an acid cleavable protecting group are provided. The labelled solid-support is reacted in a cyclical fashion to synthesize a labelled oligonucleotide on a solid-support in the 5′ to 3′ direction, having the structure: Labelled oligonucleotides are also synthesized by reacting: (i) a label reagent bearing functionality consisting of carboxylic acid, sulfonic acid, phosphonic acid, or phosphoric acid, (ii) an oligonucleotide on solid support with nucleophilic functionality, and (iii) a coupling reagent, whereby an ester, amide, thioester, sulfonamide, sulfonate, phosphonate, phosphoramidate, phosphorothioate, or phosphate bond is formed.

Abstract: The invention relates to methods for the base sequencing of deoxyribonucleic acid or ribonucleic acid, comprising the following steps: (1) immobilising DNA or RNA single strands on a planar support; (2) focussing a laser beam on a single, immobilised single strand; (3) producing DNA or RNA complimentary strand of said immobilised, focused single strand by adding a solution containing (i) at least one luminescence-tagged nucleotide on the bases of adenine, cytosine, guanine and thymine for producing a DNA or RNA complementary strand or at least one luminescence-tagged nucleotide of the bases adenine, cytosine, guanine and uracil for producing an RNA complementary strand and (ii) a polymerase, each insertion of a luminescence-tagged nucleotide into the complementary strand being detected with a single-molecule detector and the luminescence signal of the previous luminescence-tagged nucleotide being deleted before the next luminescence-tagged nucleotide is inserted.

Abstract: The invention provides a high efficiency method for combined immunocytochemistry and in situ hybridization. In one aspect, the method is used to simultaneously determining a cell phenotype and genotype by contacting a cell with an antigen-specific antibody bound to a ligand, contacting the cell with polynucleotide probe to form a complex of the probe and a nucleic acid in the cell, contacting the cell with a detectably labeled anti-ligand, and detecting the polynucleotide-probe complex and the anti-ligand-ligand complex. The presence of the anti-ligand is correlated with the presence of the antigen and the presence of the probe-nucleic acid complex is correlated with the presence of the nucleic acid in the cell.

Abstract: A system for producing substantially identical specific binding ligand (e.g., nucleic acid) probe arrays, for instance, by preparing and replicating an original master array and/or by providing a reusable assay array that is capable of being regenerated. In one embodiment the system includes the preparation and use of a) a master array surface having address sequences immobilized in the form of a patterned, and optionally random, array, b) a multi-ligand conjugate having a binding domain complementary to an address sequence, a binding domain complementary to a target sequence, and a third ligand for use in forming (e.g., by binding or polymerization) the conjugates into or onto the surface of assay array, which can be used with or upon disassociation of the address and its complementary sequences.

Abstract: The present invention relates to the discovery of novel genes encoding a fibroblast growth factor, MFGF. Therapeutics, diagnostics and screening assays based on these molecules are also disclosed.

Abstract: The invention provides a method for determining the identity of an unknown live microorganism in a mixed culture. The microorganism can be a bacterium, fungus, virus, or protozoan. The invention further provides an assay for determining the ability of a selected microorganism in a mixed culture to replicate in the presence of a chemical agent. Kits for determining the identity of a live microorganism in a mixed culture and for determining the ability of a microorganism in a mixed culture to replicate are also provided.

Abstract: A photoactivatable nucleic acid derivative composition in which one or more photoreactive group(s) are bound to a natural or synthetic nucleic acid. The photoreactive groups can be bound to the nucleic acid before, during or after its formation, and can thereafter be activated in order to attach the nucleic acid to another molecule, e.g., to the surface of a solid support. Also described is a method of preparing such a composition, and a method of using such a composition to attach the nucleic acid to a another molecule, such as that provided by the surface of a substrate used to prepare a nucleic acid chip by photolithographic techniques.

Abstract: Methods are provided for quantifying the concentration of a nucleic acid in a nucleic acid sample. The methods include contacting the nucleic acid sample with an amplifying agent, amplifying at least one predetermined locus of the nucleic acid by subjecting the sample to a number of amplification, generating an amplification curve or array, calculating the first, second or n th order derivative of the amplification curve or array, determining a maximum, minimum, or zero value of the derivative, and using the maximum, minimum, or zero value to calculate the initial concentration of the nucleic acid in the nucleic acid sample.

Abstract: The present invention relates to methods for the detection of polymorphism in polynucleotides by using hybridization of fragments of segments of a polynucleotide suspected of containing a polymorphism with an oligonucleotide having a sequence complementary to a fragment identifying the polymorphism and subsequent detection of incorporated labels in the oligonucleotide-fragment duplex.

Abstract: By immobilizing identical or different nucleic acids in a plurality of dot-like areas on a carrier which comprises a base material and compound carried on the base material, the compound having one or more alkylating groups, through intermediary of the alkylating group, a nucleic acid-immobilized substrate on which nucleic acids are firmly immobilized in fine dot area without reference to chain length of nucleic acids is provided, which enables efficiently introducing the nucleic acids onto the base material in a simple manner and can be produced with a simple apparatus.

Abstract: The sequence of a target DNA molecule is determined by preparing four chain termination reaction mixtures, one for each base type. The first set of fragments, indicative of the positions of a first type of base, are labeled with a first fluorescent label. The second set of fragments, indicative of the positions of a second type of base, are labeled with a second fluorescent label different from the first fluorescent label, The third set of fragments, indicative of the positions of a third type of base, are labeled with a third fluorescent label, different from the first and second fluorescent labels or with at least two labels, including at least one fluorescent label selected from among the first, second and third fluorescent labels.

Abstract: The present invention relates to a process for labeling a synthetic or natural ribonucleic acid (RNA). It also relates to RNA fragments, which have been labeled by fragmenting the RNA to free a terminal phosphate of each fragment for further reaction, and labeling each fragment at the freed terminal phosphate which is located at the 3′ end and/or the 5′ end of each fragment of the RNA, and to the use of such RNA fragments, for example, in the field of medical diagnosis.

Abstract: Conjugates between a minor groove binding molecule, such as the trimer of 1,2-dihydro-(3H)-pyrrolo[3,2-e]indole-7-carboxylate (CDPI3), and an oligonucleotide form unusually stable hybrids with complementary target sequences, in which the tethered CDPI3 group resides in the minor groove of the duplex. These conjugates can be used as probes and primers. Due to their unusually high binding affinity, conjugates as short as 8-mers can be used as amplification primers with high specificity and efficiency. MGB conjugation also increases the discriminatory power of short oligonucleotides, providing enhanced detection of nucleotide sequence mismatches by short oligonucleotides.

Abstract: The present invention teaches a novel approach to detecting and/or analyzing nucleic acid sequences. Generally, the invention relates to detecting and/or analyzing nucleic acid sequences using microsphere-based assays. More specifically, the invention relates to detecting and/or analyzing nucleic acid sequences using microsphere-based oligonucleotide ligation multiplexed assays. The present invention also provides methods and kits for performing microsphere-based oligonucleotide ligation assays, which include bound probes attached to addressable microspheres and free probes bearing a detectable label.

Abstract: A method for the detection of telomerase activity, for the detection of cancer cells and for the diagnosis of cancer comprising amplifying an oligonucleotide sequence extended by a DNA extension reaction with a telomerase and hybridizing the resulting amplified product with a probe labelled with a non-radioactive material to detect the telomerase activity, as well as a diagnositic kit for use in said method for the detection and diagnosis.

Abstract: Minor groove binding molecules are covalently bound to oligonucleotides which in their base sequence are complementary to a target sequence of single stranded or double stranded DNA, RNA or hybrids thereof. The covalently bound oligonucleotide minor groove binder conjugates strogly bind to the target sequence of the complementary strand.

Abstract: The present invention provides methods of preparing large polynucleotides of arbitrary sequence and in a manner that will readily lend itself to automation. The present invention provides methods of preparing a polynucleotide having at least 200 nucleotides in either a 5′ to 3′ or 3′ to 5′ direction by ligating a plurality of oligonucleotides, the assembly of which, represents the nucleotide sequence of the desired polynucleotide.

Abstract: The invention provides methods and apparatus for analyzing polynucleotide sequences by asynchronous base extension. Some applications of the invention utilize total internal reflection fluorescence microscopy to image polynucleotide molecules at single molecule resolution.

Abstract: A method for fragmenting and labeling nucleic acids is provided. The method comprises maintaining double- and single-stranded nucleic acid molecules in an aerobic or an anaerobic atmosphere, contacting the molecules with hydrogen peroxide and radical generating coordination complexes for a time and at concentrations sufficient to produce aldehyde moieties on the molecules, reacting the aldehyde moieties with amine to produce a condensation product, and labeling the condensation product.

Abstract: Oligomers which have substituents on the 2′ position are resistant to oligonucleases and furthermore can be derivatized to deliver reagents or drugs, to carry label, or to provide other properties.

Abstract: The invention provides methods and devices for analyzing sequence variations in nucleic acid samples comprising multiple loci, each having two, three or more possible allelic sequences. The method involves combining at least a first and second pair of oligonucleotide probes with the nucleic acid sample. The first pair of probes is capable of hybridizing in proximity to each other within a segment of the nucleic acid sample comprising the first locus and the second pair is capable of hybridizing in proximity to each other within a segment of the nucleic acid sample comprising the second locus. The first member of each probe pair comprises a FRET donor and the second member comprises a FRET acceptor, the FRET acceptor of the first probe pair member having a different emission spectrum from the FRET acceptor of the second probe pair.

Abstract: A method for identifying target cells and tissues which internalize known or putative ligands is provided. A ligand displaying genetic package that carries a reporter or selectable marker and presents a ligand on its surface is utilized to screen a variety of cells and tissue types for the ability to be successfully transduced by the ligand displaying genetic package.

Abstract: Compositions and methods for fluorescent detection of nucleic acids are provided. The compositions can be detected by fluorescence when hybridized to a nucleic acid containing a target sequence, but are non-fluorescent in the non-hybridized state. Alternatively, the fluorescence properties of the compositions change in a detectable manner upon hybridization to a nucleic acid containing a target sequence. Methods for synthesis and methods of use of the compositions are also provided.

Abstract: A synthetic strategy for the creation of large scale chemical diversity. Solid-phase chemistry, photolabile protecting groups, and photolithography are used to achieve light-directed spatially-addressable parallel chemical synthesis. In one particular embodiment, an array of rotated cyclic polymers is formed. In another embodiment, an array of polymers is formed based on a target polymer. The array includes systematically substituted versions of the target molecule. In another embodiment, rotated and systematically substituted cyclic polymers are formed on a substrate.

Abstract: Fluorescent, sulfonated 3,7-diamino-[8,9]benzophenoxazine dyes are provided that are especially useful for labelling biopolymers and other substrates. The dye-labelled conjugates can be used in a variety of contexts, including cell surface assays employing intact, live cells and in nucleic acid detection methods. The new dyes are water soluble and can be conjugated to a variety of substrates, such as polynucleotides, nucleosides, nucleotides, peptides, proteins, antibodies, carbohydrates, ligands, particles and surfaces.

Abstract: The present invention relates to methods for the analysis of polynucleotides including detection of variance in nucleotide sequence without the need for full sequence determination, full sequence determination of a polynucleotide, genotyping of DNA and labeling a polynucleotide fragment during the process of cleaving it into fragments.

Abstract: Nucleic acid sequences are provided that are useful as amplification primers, hybridization probes, and as a portion of molecular beacon probes for amplifying and detecting polymorphisms of the &bgr;2 adrenergic receptor gene, compositions and kits incorporating the same, and methods employing the same.

Abstract: A method for analyzing an objective substance, comprising reacting a labeled probe with an objective substance on a biological sample, said probe comprising a label substance of the formula (I): wherein A1 is an aromatic group, R1 is a hydrogen or —COCH2COCnF2n+1 and n is an integer of 1-6, which is bonded to a probe selected from the group consisting of nucleic acid, nucleic acid binding protein, low molecular ligand and receptor for ligand (except antibody) to give a fluorescent complex, reacting the complex with an objective substance on a biological sample and assaying fluorescence of the resultant fluorescent complex, a labeled nucleic acid probe and a labeled nucleotide. According to the method of the present invention, defects such as hindrance of fluorescence due to contaminant substance, low sensitivity and the like can be resolved, thereby enabling analysis on a tissue.

Abstract: The present invention provides novel methods and materials for detecting the presence of a fungus in a biological sample. The inventive methods and materials exploit the fact that the amino acid sequence of the &agr;-aminoadipate reductase molecule is highly conserved in fungi. Inventive hybridization probes, nucleic acids, PCR primers, antibodies, epitopes, reagents and methods are provided.

Abstract: The invention provides for compounds comprising nucleotide sequences 5′-P-N1-N2-G-N3-C-I-3′ and 3′-Q-N4-N5-N6-N7-N8-N9-Y-J-5′ that are the generic sequences of a mIRES of an HCV genome, and variants thereof The invention also provides assays that utilise the compounds of the invention, including assays for detection of HCV antiviral compounds.